Anti-PD-L1 antibody for detecting PD-L1

ABSTRACT

The present invention provides an anti-PD-L1 antibody capable of staining tumor cells such as melanoma cells. 
     An anti-PD-L1 antibody comprising (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDP (SEQ ID NO: 5). A composition for detecting PD-L1, comprising the above antibody as an active ingredient. A method for preparing the above antibody is also provided.

CLAIM FOR PRIORITY

This application is a continuation of U.S. application Ser. No. 16/491,145, filed Sep. 4, 2019 which is a U.S. National Stage Filing under 35 U.S.C. 371 from International Application No. PCT/JP2018/011895, filed on Mar. 23, 2018, and published as WO2018/181064 on Oct. 4, 2018, which claims the benefit of priority to Japanese Application No. 2017-061389, filed on Mar. 27, 2017; the benefit of priority of each of which is hereby claimed herein, and which applications and publication are hereby incorporated herein by reference in their entirety.

TECHNICAL FIELD

The present invention relates to an anti-PD-L1 antibody for detecting PD-L1.

BACKGROUND ART

Malignant melanoma originating from melanocytes is one of the most commonly observed malignant tumors in the canine oral cavity (Non-Patent Document No. 1: Todoroff et al., J Am Vet Med Assoc. 1979 Sep. 15; 175(6):567-71). Since this type of melanoma generally tends to be highly invasive and metastatic, early diagnosis and treatment are desired. On the other hand, malignant melanoma has a wide tissue variation, presenting various morphologies such as epithelial-like, round cell-like or fibrosarcoma-like morphology. Thus, malignant melanoma is one of those tumors which involve difficulty in tissue diagnosis. Although confirmation of melanin pigment is important for their diagnosis, a large number of malignant melanomas do not have melanin pigment and, sometimes, diagnosis cannot be made with histological observations alone. This has led to searches for diagnostic markers that can be used in immunohistochemical techniques. Among such markers, Melan A/MART-1, vimentin, S100, neuron-specific enolase and the like have been reported to be useful (Non-Patent Document No. 2: Ramos-Vara et al., Vet Pathol. 2000 November; 37(6):597-608). However, even Melan A/MART-1, the most widely used diagnostic marker, has a positive rate not higher than about 60% which varies among reports (Non-Patent Document No. 3: Koenig et al., Vet Pathol. 2001 July; 38(4):427-35). Because of this sensitivity problem, the utility of Melan A/MART-1 in actual diagnosis is still arguable. Further, Melan A/MART-1 is not stained in amelanotic melanoma (Non-Patent Document No. 3: Koenig et al., Vet Pathol 2001 July 38(4):427-35), so its application to diagnosis is limited. Under these circumstances, it is desired to develop highly sensitive, novel diagnostic markers to malignant melanoma.

PRIOR ART LITERATURE Non-Patent Documents

-   Non-Patent Document No. 1: Todoroff et al., J Am Vet Med Assoc. 1979     Sep. 15, 175(6):567-71 -   Non-Patent Document No. 2: Ramos-Vara et al., Vet Pathol. 2000     November; 37(6):597-608 -   Non-Patent Document No. 3: Koenig et al., Vet Pathol. 2001 July;     38(4):427-35

DISCLOSURE OF THE INVENTION Problem for Solution by the Invention

It is an object of the present invention to provide a PD-L1 antibody capable of staining tumor cells such as melanoma cells.

Means to Solve the Problem

The present inventors have established a number of monoclonal antibodies which react with the PD-L1 protein of various animals. It has been revealed that, among those monoclonal antibodies, a rat anti-bovine PD-L1 monoclonal antibody (6C11-A11) is capable of staining melanoma tumor cells very strongly. Currently, this monoclonal antibody is used for selecting candidate dogs for therapy with chimeric antibodies. The subject PD-L1 antibody (6C11-3A11) is also capable of immunohistochemically staining ovine, porcine and bovine PD-L1 proteins. Further, the present inventors have determined the CDRs (complementarity-determining regions) of the variable regions of the subject PD-L1 antibody (6C11-3A11). The present invention has been achieved based on these findings.

A summary of the present invention is as described below.

(1) An anti-PD-L1 antibody comprising (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the ammo acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the ammo arid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5). (2) The antibody of (1) above, which is derived from rat. (3) The antibody of (2) above, which is a rat anti-bovine PD-L1 antibody. (4) The antibody of (3) above, wherein the light chain variable region has the amino acid sequence as shown in SEQ ID NO: 6 and the heavy chain variable region has the amino acid sequence as shown m SEQ ID NO: 7. (5) The antibody of any one of (1) to (4) above, wherein die light chain constant region has the amino acid sequence of the constant region of kappa chain. (6) The antibody of any one of (1) to (5) above, wherein the heavy chain constant region has the amino acid sequence of the constant region of IgG2a. (7) The antibody of (5) or (6) above, wherein the light chain constant region has the amino acid sequence as shown in any one of SEQ ID NOS: 8, 10 to 12 and the heavy chain constant region has the amino acid sequence as shown in SEQ ID NO: 9 or 13. (8) The antibody of any one of (1) to (7) above which has a four-chain structure comprising two light chains and two heavy chains. (9) A composition for detecting PD-L1, comprising the antibody of any one of (1) to (8) above as an active ingredient. (10) The composition of (9) above for use in diagnosis of cancers and/or inflammations. (11) The composition of (10) above, wherein the cancers and/or inflammations are selected from the group consisting of neoplastic diseases, leukemia, Johne's disease, anaplasmosis, bacterial mastitis, mycotic mastitis, mycoplasma infections (such as mycoplasma mastitis, mycoplasma pneumonia or the like), tuberculosis, Theileria orientalis infection, cryptosporidiosis, coccidiosis, trypanosomiasis and leishmaniasis. (12) The composition of (9) above for use in selecting subject animals suitable for therapy with anti-PD-L1 antibodies. (13) A DNA encoding the anti-PD-L1 antibody of (1) above. (14) A vector comprising the DNA of (13) above. (15) A host cell transformed with the vector of (14) above. (16) A method of preparing an antibody, comprising culturing the host cell of (15) above and collecting an anti-PD-L1 antibody from the resultant culture. (17) A DNA encoding the light chain of an anti-PD-L1 antibody, said light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS arid CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO. 2). (18) A DMA encoding the heavy chain of an anti-PD-L1 antibody, said heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5).

Effect of the Invention

According to the present invention, a novel anti-PD-L1 anti-body capable of staining tumor cells, such as melanoma cells, has been obtained.

The present specification encompasses the contents disclosed in the specification and/or the drawings of Japanese Patent Application No. 2017-61389 based on which the present patent application claims priority.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 Binding specificity of rat anti-bovine PD-L1 antibody 6C11-3A11. Rat anti-bovine PD-L1 antibody 6C11-3A11 did not bind to EGFP expressing cells, but specifically bound to canine PD-L1-EGFP expressing cells.

FIG. 2 Predicted CDR regions of rat anti-bovine PD-L1 antibody 6C11-3A11. The regions of CDR1, CDR2 and CDR3 in the light chain variable region and the heavy chain variable region of rat anti-bovine PD-L1 antibody 6C11-3A11 are shown.

FIG. 3 Comparative immunohistochemical staining images of canine melanoma. Left: stained with a commercial antibody (MelanA antibody). Tumor cells were stained very weakly. Right stained with the PD-L1 antibody 6C11-3A11 established by the present inventors. Tumor cells were stained very strongly.

FIG. 4 Immunohistochemical staining image of canine melanoma.

FIG. 5-1 Immunohistochemical staining images of other tumors. Upper left: case of canine lymphoma. Upper right: case of canine osteosarcoma. Lower left: case 1 of canine renal cell carcinoma. Lower right: case 2 of canine renal cell carcinoma.

FIG. 5-2 Immunohistochemical staining images of other tumors. Left: Case of canine squamous cell carcinoma. Right: case of canine fibrosarcoma.

FIG. 6 Immunohistochemical staining image of a case of ovine listeriosis. Left: PD-L1 staining image of a brain lesion of ovine listeriosia exhibiting neurologic symptoms. Right: enlarged photograph of the left image.

FIG. 7 Immunohistochemical staining images of porcine infections. Left: case of porcine circovirus type 2 infection. Right: case of porcine mycoplasma pneumonia.

FIG. 8 Alignment of amino acid sequences of the constant region of rat Ig kappa chain (light chain).

FIG. 9 Alignment of amino acid sequences of the constant region of rat IgG2a chain (heavy chain).

FIG. 10 Schematic drawing of pDC6 vector and a rat-human chimeric anti-PD-L1 antibody.

FIG. 11 Binding of rat anti-bovine PD-L1 antibodies 6C11-3A11 and 6G7-E1 to canine PD-L1-EGFP expressing cells. 6C11-3A11 specifically bound to canine PD-L1-EGFP expressing cells.

FIG. 12 Immunohistochemical staining images of skin squamous carcinoma, nasal adenocarcinoma and transitional cell carcinoma in dogs. No specific signals were detected with 6G7-E1. Tumor cells were stained 6C11-3A11.

FIG. 13 Immunohistochemical staining images of anal sac gland carcinoma, soft tissue sarcoma and osteosarcoma in dogs. In anal sac gland carcinoma and soft tissue sarcoma, no specific signals were detected with 6G7-E1, but tumor cells were stained with 6C11-3A11. In osteosarcoma, both antibodies stained tumor cells, but stronger signals were obtained with 6C11-3A11.

FIG. 14 Immunohistochemical staining images of oral malignant melanoma, mammary adenocarcinoma histiocytic sarcoma, diffuse large B-cell lymphoma and transmissible venereal tumor in dogs using 6C11-3A11. In the tumor species other than transmissible venereal tumor, PD-L1 on tumor cells was stained.

FIG. 15 Binding of rat anti-bovine PD-L1 antibody 6C11-3A11 to bovine PD-L1-EGFP expressing cells. 6C11-3A11 specifically bound to bovine PD-L1-EGFP expressing cells.

FIG. 16 Immunohistochemical staining images of ileal lesions of cattle naturally and experimentally infected with Mycobacterium avium subsp. paratuberculosis, using (a) 6C11-3A11 and (b) Ziehl-Neelsen staining. 6C11-3A11 detected PD-L1 expression in cells infected with M. avium subsp. paratuberculosis (positive in Ziehl-Neelsen staining).

BEST MODES FOR CARRYING OUT THE INVENTION

Hereinbelow, the present invention will be described in detail.

The present invention provides an anti-PD-L1 antibody comprising (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5).

CDR1, CDR2 and CDR3 in the light chain variable region (VL) of rat anti-bovine PD-L1 antibody 6C11-3A11 (monoclonal antibody) established by the present inventors are a region consisting of the amino acid sequence of KSISKY (SEQ ID NO: 1), a region consisting of the amino acid sequence of SGS and a region consisting of the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2), respectively (see FIG. 2 ).

Further, CDR1, CDR2 and CDR3 in the heavy chain variable region (VH) of rat anti-bovine PD-L1 antibody 6C11-3A11 are a region consisting of the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), a region consisting of the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and a region consisting of the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5), respectively (see FIG. 2 ).

In the ammo acid sequences of KSISKY (SEQ ID NO: 1), SGS and QQHNEYPLT (SEQ ID NO: 2), as well as the amino acid sequences of GYTFTDYI (SEQ ID NO: 3), INPDSGGN (SEQ ID NO: 4) and ARGITMMVVISHWKFDF (SEQ ID NO: 5), one, two, three, four or five amino acids may be deleted, substituted or added. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as CDR of VL or CDR of VH of the PD-L1 antibody.

As used herein, the term “antibody” is a concept encompassing not only full-length antibodies but also antibodies of smaller molecular sizes such as Fab, F(ab)′₂, ScFv, Diabody, V_(H), V_(L), Sc(Fv)₂, Bispecific sc(Fv)₂, Minibody, scFv-Fc monomer and scFv-Fc dimer.

The anti-PD-L1 antibody of the present invention may be derived from rat. For example, the anti-PD-L1 antibody may be a rat anti-bovine PD-L1 antibody.

The amino acid sequence of the VL and the amino acid sequence of the VH of rat anti-bovine PD-L1 antibody 6C11-3A11 (monoclonal antibody) are shown in SEQ ID NOS: 6 and 7, respectively. The amino acid sequences as shown in SEQ ID NOS: 6 and 7 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) amino acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as VL or VH of the PD-L1 antibody.

There are two types of immunoglobulin light chain, which are called Kappa chain (κ) and Lambda chain (λ). In the anti-PD-L1 antibody of the present invention, the light chain constant region (CL) may have the amino acid sequence of the constant region of either Kappa chain or Lambda chain. However, the relative abundance of Lambda chain is higher in ovine, feline, canine and equine, and that of Kappa chain is higher in mouse, rat, human and porcine. Rat anti-bovine PD-L1 antibody 6C11-3A11 (monoclonal antibody) is a rat-derived IgG2a, and the CL thereof has the amino acid sequence of the constant region of Kappa chain.

The heavy chain constant region (CH) of the anti-PD-L1 antibody of the present invention may have the amino acid sequence of the constant region of rat IgG2a. Immunoglobulin heavy chain is classified into γ chain, μ chain, α chain, δ chain and ε chain depending on the difference in constant region. According to the type of heavy chain present, five classes (isotypes) of immunoglobulin are formed: they are IgG, IgM, IgA, IgD and IgE.

Immunoglobulin G (IgG) accounts for 70-75% of human immunoglobulins and is the most abundantly found monomeric antibody in plasma. IgG has a four-chain structure consisting of two light chains and two heavy chains. Human IgG1, IgG2 and IgG4 have molecular weights of about 146,000, whereas human IgG3 has a long hinge region that connects Fab region and Fc region and has a larger molecular weight of 170,000. Human IgG1 accounts for about 65%, human IgG2 about 25%, human IgG3 about 7%, and human IgG4 about 3% of human IgG. They are uniformly distributed inside and outside of blood vessels. Having a strong affinity for Fc receptors and complement factors on effector cell surfaces, human IgG1 induces antibody-dependent cell cytotoxicity (ADCC) and also activates complements to induce complement-dependent cell cytotoxicity (CDC). Human IgG2 and IgG4 are low at ADCC and CDC activities because their affinity for Fc receptors and complement factors is low.

Immunoglobulin M (IgM), which accounts for about 10% of human immunoglobulins, is a pentameric antibody consisting of five basic four-chain structures joined together. It has a molecular weight of 970,000. Usually occurring only in blood, IgM is produced against infectious microorganisms and takes charge of early stage immunity.

Immunoglobulin A (IgA) accounts for 10-15% of human immunoglobulins. It has a molecular weight of 160,000. Secreted IgA is a dimeric antibody consisting of two IgA molecules joined together. IgA1 is found in serum, nasal discharge, saliva and breast milk. In intestinal juice, IgA2 is found abundantly.

Immunoglobulin D (IgD) is a monomeric antibody accounting for no more than 1% of human immunoglobulins. IgD is found on B cell surfaces and involved in induction of antibody production.

Immunoglobulin E (IgE) is a monomeric antibody that occurs in an extremely small amount, accounting for only 0.001% or less of human immunoglobulins. Immunoglobulin E is considered to be involved in immune response to parasites but in advanced countries where parasites are rare, IgE is largely involved in bronchial asthma and allergy among other things.

With respect to rat, sequences of IgG1, IgG2a, IgG2b and IgG2c have been identified as the heavy chain of IgG. Rat anti-bovine PD-L1 antibody 6C11-3A11 has the amino acid sequence of the CH of IgG2a.

In the antibody of the present invention, it is more preferable that the CL has the amino acid sequence of the constant region of Kappa chain and that the CH has the amino acid sequence of the constant region of IgG2a.

The amino acid sequence and the nucleotide sequence of the VL of rat anti-bovine PD-L1 antibody 6C11-3A11 identified by the present inventors are shown in SEQ ID NOS: 6 and 14, respectively.

The amino acid sequence and the nucleotide sequence of the VH of rat anti-bovine PD-L1 antibody 6C11-3A11 identified by the present inventors are shown in SEQ ID NOS: 7 and 15, respectively.

The amino acid sequence and the nucleotide sequence of the CL (Kappa chain) of rat anti-bovine PD-L1 antibody 6C11-3A11 identified by the present inventors are shown in SEQ 3 ID NOS: 8 and 16, respectively. These sequences are identical with the sequences registered at GenBank (a nucleotide sequence database provided by National Center for Biotechnology Information (NCBI)) under accession numbers #XM_08775358.2, #BC062802.1, #BC088255.1, #L22653.1, #L22655.1 and #M14434.1.

The amino acid sequence and the nucleotide sequence of the CH (IgG2a) of rat anti-bovine PD-L1 antibody 6C11-3A11 identified by the present inventors are shown in SEQ ID NOS: 9 and 17, respectively. These sequences are identical with the sequences registered at GenBank under accession numbers #BC088240.1, #BC091257.1, 1909 BC09I272.1, #BC088423.1, #L22652.1 and #L22654.1.

Amino acid sequences and nucleotide sequences of CLs and CHs for rat antibodies other than the above may be obtained from known databases for use in the present invention.

As an amino acid sequence and a nucleotide sequence of rat Ig Kappa chain, the sequence registered at GenBank under accession number #V01241.1 is shown in SEQ ID NOS: 10 and 18.

As an amino acid sequence and a nucleotide sequence of rat Ig Kappa chain, the sequence registered at GenBank under accession number #X16129.1 is shown in SEQ ID NOS: 11 and 19.

As an amino acid sequence and a nucleotide sequence of rat Ig Kappa chain, the sequence registered at GenBank under accession number #DQ402471.1 is shown in SEQ ID NOS: 12 and 20.

As the CH of rat IgG2a, the sequence registered at GenBank under accession number #DQ402472.1 is shown in SEQ ID NOS: 13 and 21.

The anti-PD-L1 antibody of the present invention may be an anti-PD-L1 antibody in which the CL has the amino acid sequence as shown in any one of SEQ ID NOS: 8 and 10 to 12 and the CH has the amino acid sequence as shown in SEQ ID NO: 9 or 13.

The amino acid sequences as shown in SEQ ID NOS: 8 to 13 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) ammo acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as CL or CH of the PD-L1 antibody.

Alignments of amino acid sequences of the CL and the CH of a rat anti-PD-L1 antibody are shown in FIG. 8 and FIG. 9 , respectively. The above-described mutations such as deletion, substitution or addition of amino acids may suitably have occurred at the mutation sites as shown in FIGS. 8 and 9 or at the vicinity thereof.

The anti-PD-L1 antibody of the present invention may be a chimeric antibody. The VL and the VH of the antibody may be suitable derived from rat. For example, the VL may be the VL of a rat anti-PD-L1 antibody (e.g., 6C11-3A11); the VH may be the VH of a rat anti-PD-L1 antibody; and the CL and the CH may be derived from an animal other than rat. For example, when a rat antibody a chimerized using the constant regions of a mouse antibody, the resulting chimeric antibody will be useful for testing and diagnosis because various secondary antibodies to mouse antibodies are commercially available. Amino acid sequences and nucleotide sequences of the CLs and the CHs of antibodies of animals other than rat may be obtained from known databases for use in the present invention.

Amino acid sequences and nucleotide sequences of CLs and CHs for human, mouse, bovine, canine, ovine, porcine and water buffalo are summarized in the table below.

TABLE GenBank Acces- Nucleotide Amino Acid sion IMGT Refer- Species Ig Domain Sequence Sequence No. Database ence Human Human IgG GAGTCCAAATATGGT ESKYGPPCPSCPAPEFL K01316 http:// Ellison  (Scien- Ig 4 CCCCCATGCCCATCA GGPSVFLFPPKPKDTL www. J. et tific heavy vari- TGCCCAGCACCTGA MISRTPEVTCVVVDVSQ imgt.org/ al., Name: chain ant 1 GTTCCTGGGGGGAC EDPEVQFNWYVDGVEV IMGT DNA, 1, Homo con- CATCAGTCTTCCTGT HNAKTKPREEQFNSTY reper 11-18 sapiens) stant TCCCCCCAAAACCCA RVVSVLTVLHQDWLNG toire/ (1981). region AGGACACTCTCATGA KEYKCKVSNKGLPSSIE index.php? PMID: (CH1~ TCTCCCGGACCCCTG KTISKAKGQPREPQVYT section = 6299662 CH3) AGGTCACGTGCGTG LPPSQEEMTKNQVSLT Locus GTGGTGGACGTGAG CLVKGFYPSDIAVEWES Genes & CCAGGAAGACCCCG NGQPENNYKTTPPVLD reper AGGTCCAGTTCAACT SDGSFFLYSRLTVDKSR toire = GGTACGTGGATGGC WQEGNVFSCSVMHEAL gene GTGGAGGTGCATAA HNHYTQKSLSLSLGK* table & TGCCAAGACAAAGC (SEQ ID NO: 30) species = CGCGGGAGGAGCAG human & TTCAACAGCACGTAC group = CGTGTGGTCAGCGT IGHC CCTCACCGTCCTGCA CCAGGACTGGCTGA ACGGCAAGGAGTAC AAGTGCAAGGTCTC CAACAAAGGCCTCC CGTCCTCCATCGAGA AAACCATCTCCAAAG CCAAAGGGCAGCCC CGAGAGCCACAGGT GTACACCCTGCCCCC ATCCCAGGAGGAGA TGACCAAGAACCAG GTCAGCCTGACCTG CCTGGTCAAAGGCTT CTACCCCAGCGACAT CGCCGTGGAGTGGG AGAGCAATGGGCAG CCGGAGAACAACTA CAAGACCACGCCTC CCGTGCTGGACTCC GACGGCTCCTTCTTC CTCTACAGCAGGCTA ACCGTGGACAAGAG CAGGTGGCAGGAGG GGAATGTCTTCTCAT GCTCCGTGATGCAT GAGGCTCTGCACAA CCACTACACACAGAA GAGCCTCTCCCTGTC TCTGGGTAAATGA (SEQ ID NO: 31) IgG GAGTCCAAATATGGT ESKYGPPCPSCPAPEFL AJ001563 Brusco  4 CCCCCGTGCCCATCA GGPSVFLFPPKPKDTL A,  vari- TGCCCAGCACCTGA MISRTPEVTCVVVDVSQ et al., ant 2 GTTCCTGGGGGGAC EDPEVQFNWYVDGVEV Eur. J. CATCAGTCTTCCTGT HNAKTKPREEQFNSTY Immuno-  TCCCCCCAAAACCCA RVVSVLTVVHQDWLNG gen AGGACACTCTCATGA KEYKCKVSNKGLPSSIE et., TCTCCCGGACCCCTG KTISKAKGQPREPQVYT 25, AGGTCACGTGCGTG LPPSQEEMTKNQVSLT 349-355 GTGGTGGACGTGAG CLVKGFYPSDIAVEWES (1998). CCAGGAAGACCCCG NGQPENNYKTTPPVLD PMID: AGGTCCAGTTCAACT SDGSFFLYSRLTVDKSR 9805657 GGTACGTGGATGGC WQEGNVFSCSVMHEAL GTGGAGGTGCATAA HNHYTQKSLSLSLGK* TGCCAAGACAAAGC (SEQ ID NO: 32) CGCGGGAGGAGCAG TTCAACAGCACGTAC CGTGTGGTCAGCGT CCTCACCGTCGTGCA CCAGGACTGGCTGA ACGGCAAGGAGTAC AAGTGCAAGGTCTC CAACAAAGGCCTCC CGTCCTCCATCGAGA AAACCATCTCCAAAG CCAAAGGGCAGCCC CGAGAGCCACAGGT GTACACCCTGCCCCC ATCCCAGGAGGAGA TGACCAAGAACCAG GTCAGCCTGACCTG CCTGGTCAAAGGCTT CTACCCCAGCGACAT CGCCGTGGAGTGGG AGAGCAATGGGCAG CCGGAGAACAACTA CAAGACCACGCCTC CCGTGCTGGACTCC GACGGCTCCTTCTTC CTCTACAGCAGGCTA ACCGTGGACAAGAG CAGGTGGCAGGAGG GGAATGTCTTCTCAT GCTCCGTGATGCAT GAGGCTCTGCACAA CCACTACACGCAGA AGAGCCTCTCCCTGT CTCTGGGTAAATGA (SEQ ID NO: 33) IgG GCACCTGAGTTCCTG APEFLGGPSVFLFPPKP AJ001564 4 GGGGGACCATCAGT KDTLMISRTPEVTCVVV vari- CTTCCTGTTCCCCCC DVSQEDPEVQFNWYVD ant 3 AAAACCCAAGGACA GVEVHNAKTKPREEQF CTCTCATGATCTCCC NSTYRVVSVLTVLHQD GGACCCCTGAGGTC WLNGKEYKCKVSNKG ACGTGCGTGGTGGT LPSSIEKTISKAKGQPR GGACGTGAGCCAGG EPQVYTLPPSQEEMTK AAGACCCCGAGGTC NQVSLTCLVKGFYPSDI CAGTTCAACTGGTAC AVEWESNGQPENNYKT GTGGATGGCGTGGA TPPVLDSDGSFFLYSKL GGTGCATAATGCCA TVDRSRWQEGNVFSCS AGACAAAGCCGCGG VMHEALHNHYTQKSLS GAGGAGCAGTTCAA LSLGK* CAGCACGTACCGTG (SEQ ID NO: 34) TGGTCAGCGTCCTCA CCGTCCTGCACCAG GACTGGCTGAACGG CAAGGAGTACAAGT GCAAGGTCTCCAAC AAAGGCCTCCCGTC CTCCATCGAGAAAAC CATCTCCAAAGCCAA AGGGCAGCCCCGAG AGCCACAGGTGTAC ACCCTGCCCCCATCC CAGGAGGAGATGAC CAAGAACCAGGTCA GCCTGACCTGCCTG GTCAAAGGCTTCTAC CCCAGCGACATCGC CGTGGAGTGGGAGA GCAATGGGCAGCCG GAGAACAACTACAA GACCACGCCTCCCG TGCTGGACTCCGAC GGCTCCTTCTTCCTC TACAGCAAGCTCACC GTGGACAAGAGCAG GTGGCAGGAGGGGA ACGTCTTCTCATGCT CCGTGATGCATGAG GCTCTGCACAACCAC TACACGCAGAAGAG CCTCTCCCTGTCTCT GGGTAAATGA (SEQ ID NO: 35) Human Ig ACTGTGGCTGCACG TVAAPSVFIFPPSDEQL X96754 http:// Ig  kappa ATCTGTCTTCATCTT KSGTASVVGLLNNFYPR www. light (CK) CCCGCCATCTGATGA EAKVQWKVDNALQSG imgt.org/ chain GCAGTTGAAATCTG NSQESVTEQDSKDSTYS IMGT con- GAACTGCCTCTGTTG LSSTLTLSKADYEKHKV reper stant TGTGCCTGCTGAATA YACEVTHQGLSSPVTKS toire/ region ACTTCTATCCCAGAG FNRGEC* index.php? AGGCCAAAGTACAG (SEQ ID NO: 28) section = TGGAAGGTGGATAA Locus CGCCCTCCAATCGG Genes & GTAACTCCCAGGAG reper AGTGTCACAGAGCA toire = GGACAGCAAGGACA gene GCACCTACAGCCTCA table & GCAGCACCCTGACG species = CTGAGCAAAGCAGA human & CTACGAGAAACACA group = AAGTCTACGCCTGC IGKC GAAGTCACCCATCA GGGCCTGAGCTCGC CCGTCACAAAGAGC TTCAACAGGGGAGA GTGTTAG (SEQ ID NO: 29) Mouse Mouse IgG1 GCCAAAACGACACCCCCA AKTTPPSVYPLAPGSAAQ J00453 http:// Honjo T. (Scien- Ig vari- TCTGTCTATCCACTGGCC TNSMVTLGCLVKGYFPE AH00530 www. et al., tific heavy ant 1 CCTGGATCTGCTGCCCAA PVTVTWNSGSLSSGVHT 9  imgt.org/ Cell,  Name: chain ACTAACTCCATGGTGACC FPAVLESDLYTLSSSVTV V00793 IMGT 18, Mus con- CTGGGATGCCTGGTCAAG PSSPRPSETVTCNVAHPA D78344 reper 559-568 muscu- stant GGCTATTTCCCTGAGCCA SSTKVDKKIVPRDCGCKP toire/ (1979). lus) region GTGACAGTGACCTGGAAC CICTVPEVSSVFIFPPKP index.php? PMID: (CH1~ TCTGGATCCCTGTCCAGC KDVLTITLTPKVTCVVVD section = 115593 CH3) GGTGTGCACACCTTCCCA ISKDDPEVQFSWFVDDVE Locus Akahori GCTGTCCTGGAGTCTGAC VHTAQTQPREEQFNSTFR Genes & Y. and CTCTACACTCTGAGCAGC SVSELPIMHQDWLNGKE reper Kurosawa TCAGTGACTGTCCCCTCC FRCRVNSAAFPAPIEKTI toire = Y., AGCCCTCGGCCCAGCGA SKTKGRPKAPQVYTIPPP gene Geno- GACCGTCACCTGCAACGT KEQMAKDKVSLTCMITD table & mics., TGCCCACCCGGCCAGCAG FFPEDITVEWQWNGQPA species = 41, CACCAAGGTGGACAAGAA ENYKNTQPIMNTNGSYF Mus_ 100-104 AATTGTGCCCAGGGATTG VYSKLNVQKSNWEAGNT musculus & (1997). TGGTTGTAAGCCTTGCAT FTCSVLHEGLHNHHTEK group = PMID: ATGTACAGTCCCAGAAGT SLSHSPGK IGHC 9126488 ATCATCTGTCTTCATCTT (SEQ ID NO: 44) CCCCCCAAAGCCCAAGGA TGTGCTCACCATTACTCT GACTCCTAAGGTCACGTG TGTTGTGGTAGACATCAG CAAGGATGATCCCGAGGT CCAGTTCAGCTGGTTTGT AGATGATGTGGAGGTGCA CACAGCTCAGACGCAACC CCGGGAGGAGCAGTTCA ACAGCACTTTCCGCTCAG TCAGTGAACTTCCCATCA TGCACCAGGACTGGCTCA ATGGCAAGGAGTTCAAAT GCAGGGTCAACAGTGCA GCTTTCCCTGCCCCCATC GAGAAAACCATCTCCAAA ACCAAAGGCAGACCGAA GGCTCCACAGGTGTACAC CATTCCACCTCCCAAGGA GCAGATGGCCAAGGATAA AGTCAGTCTGACCTGCAT GATAACAGACTTCTTCCC TGAAGACATTACTGTGGA GTGGCAGTGGAATGGGC AGCCAGCGGAGAACTACA AGAACACTCAGCCCATCA TGAACACGAATGGCTCTT ACTTCGTCTACAGCAAGC TCAATGTGCAGAAGAGCA ACTGGGAGGCAGGAAAT ACTTTCACCTGCTCTGTG TTACATGAGGGCCTGCAC AACCACCATACTGAGAAG AGCCTCTCCCACTCTCCT GGTAAATGA (SEQ ID NO: 45) IgG1 GCCAAAACGACACCCCCA AKTTPPSVYPLAPGSAAQ L35252 Honjo T. vari- TCTGTCTATCCACTGGCC TNSMVTLGCLVKGYFPE et al., ant 2 CCTGGATCTGCTGCCCAA PVTVTWNSGSLSSGVHT Cell,  ACTAACTCCATGGTGACC FPAVLQSDLYTLSSSVTV 18, CTGGGATGCCTGGTCAAG PSSTWPSQTVTCNVAHP 559-568 GGCTATTTCCCTGAGCCA ASSTKAMKKIVPRDCGCK (1979). GTGACAGTGACCTGGAAC PCICTVPEVSSVFIFPPKP PMID: TCTGGATCCCTGTCCAGC KDVLTITLTPKVTCVVVD 115593 GGTGTGCACACCTTCCCA ISKDDPEVQFSWFVDDV GCTGTCCTGCAGTCTGAC EVHTAQTKPREEQINSTF CTCTACACTCTGAGCAGC RSVSELPIMHQDWLNGK TCAGTGACTGTCCCCTCC EFKCRVNSAAFPAPIEKT AGCACCTGGCCCAGCCAG ISKTKGRPKAPQVYTIPP ACCGTCACCTGCAACGTT PKEQMAKDKVSLTCMIT GCCCACCCGGCCAGCAG NFFPEDITVEWQWNGQP CACCAAGGTGGACAAGAA AENYKNTQPIMDTDGSY AATTGTGCCCAGGGATTG FVYSKLNVQKSNWEAGN TGGTTGTAAGCCTTGCAT TFTCSVEHEGLHNHHTE ATGTACAGTCCCAGAAGT KSLSHSPGK ATCATCTGTCTTCATCTT (SEQ ID NO: 46) CCCCCCAAAGCCCAAGGA TGTGCTCACCATTACTCT GACTCCTAAGGTCACGTG TGTTGTGGTAGACATCAG CAAGGATGATCCCGAGGT CCAGTTCAGCTGGTTTGT AGATGATGTGGAGGTGCA CACAGCTCAGACGAAACC CCGGGAGGAGCAGATCA ACAGCACTTTCCGTTCAG TCAGTGAACTTCCCATCA TGCACCAGGACTGGCTGA ATGGCAAGGAGTTCAAAT GCAGGGTCAACAGTGCA GCTTTCCCTGCCCCCATC GAGAAAACCATCTCCAAA ACCAAAGGCAGACCGAA GGCTCCACAGGTGTACAC CATTCCACCTCCCAAGGA GCAGATGGCCAAGGATAA AGTCAGTCTGACCTGCAT GATAACAAACTTCTTCCC TGAAGACATTACTGTGGA GTGGCAGTGGAATGGGC AGCCAGCGGAGAACTACA AGAACACTCAGCCCATCA TGGACACAGATGGCTCTT ACTTCGTCTACAGCAAGC TCAATGTGCAGAAGAGCA ACTGGGAGGCAGGAAAT ACTTTCACCTGCTCTGTG TTACATGAGGGCCTGCAC AACCACCATACTGAGAAG AGCCTCTCCCACTCTCCT GGTAAATGA (SEQ ID NO: 47) IgG2a GCCAAAACAACAGCCCCA AKTTAPSVYPLAPVCGDT J00470 Yamawak vari- TCGGTCTATCCACTGGCC TGSSVTLGCLVKGYFPEP AH00530 i- ant 1 CCTGTGTGTGGAGATACA VTLTWNSGSLSSGVHTFP 9  Kataoka  ACTGGCTCCTCGGTGACT AVLQSDLYTLSSSVTVTS V00825 Y. CTAGGATGCCTGGTCAAG STWPSQSITCNVAHPASS V00766 et al., GGTTATTTCCCTGAGCCA TKVDKKIEPRGPTIKPCP D78344 Nucleic GTGACCTTGACCTGGAAC PCKCPAPNLLGGPSVFIF Acids  TCTGGATCCCTGTCCAGT PPKIKDVLMISLSPIVTC Res., 9, GGTGTGCACACCTTCCCA VVVDVSEDDPDVQISWFV 1365- GCTGTCCTGCAGTCTGAC NNVEVHTAQTQTHREDY 1381 CTCTACACCCTCAGCAGC NSTLRVVSALPIQHQDW (1981).  TCAGTGACTGTAACCTCG MSGKEFKCKVNNKDLPA PMID:  AGCACCTGGCCCAGCCAG PIERTISKPKGSVRAPQV 6262729 TCCATCACCTGCAATGTG YVLPPPEEEMTKKQVTL Ollo R.  GCCGACCCGGCAAGCAG TCMVTDFMPEDIYVEWT et al., CACCAAGGTGGACAAGAA NNGKTELNYKNTEPVLD Proc. AATTGAGCCCAGAGGGGC SDGSYFMYSKLRVEKKN Natl. CACAATCAAGCCCTGTCC WVERNSYSCSVVHEGLH Acad. TCCATGCAAATGCCCAGC NHHTTKSFSRTPGK Sci. ACCTAACCTCTTGGGTGG (SEQ ID NO: 48) U.S.A., ACCATCCGTCTTCATCTT 78, CCCTCCAAAGATCAAGGA 2442- TGTACTCATGATCTCCCT 2446  GAGCCCCATAGTCACATG (1981). TGTGGTGGTGGATGTGAG PMID: CGAGGATGACCCAGATGT 6787604 CGAGATCAGCTGGTTTGT Sikorav GAACAACGTGGAAGTACA J. L. CACAGCTCAGACACAAAC et al., CCATAGAGAGGATTACAA Nucleic CAGTACTCTCCGGGTGGT Acids CAGTGCCCTCCCCATCCA Res., 8, GCACCAGGACTGGATGA 3143- GTGGCAAGGAGTTCAAAT 3155 GCAAGGTCAACAACAAAG (1980). ACCTCCGAGCGCCCATCG PMID: AGAGAACCATCTCAAAAC 6777755 CCAAAGGGTCAGTAAGAG Akahori CTCCACAGGTATATGTCT Y., and TGCCTCCACCAGAAGAAG Kurosawa AGATGACTAAGAAACAGG Y., TCACTCTGACCTGCATGG Geno- TCACAGACTTCATGCCTG mics., AAGACATTTACGTGGAGT 41, GGACCAACAACGGGAAA 100-104 ACAGAGCTAAACTACAAG (1997). AACACTGAACCAGTCCTG PMID: GACTCTGATGGTTCTTAC 9126488 TTCATGTACAGCAAGCTG AGAGTGGAAAAGAAGAA CTGGGTGGAAAGAAATAG CTACTCCTGTTCAGTGGT CCACGAGGGTCTGCACAA TCACCACACGACTAAGAG CTTCTCCCGGACTCCGGG TAAATGA (SEQ ID NO: 49) IgG2a GCCAAAACAACAGCCCCA AKTTAPSYYPLAPVCGDT X16997 Morgado vari- TCGGTCTATCCACTGGCC TGSSVTLGCLVKGYFPEP M. G. et ant 2 CCTGTGTGTGGAGATACA VTLTWNSGSLSSGVHTFP al., ACTGGCTCCTCGGTGACT AVLQSDLYTLSSSVTVTS EMBO J., CTAGGATGCCTGGTCAAG STWPSQSITCNVAHPASS 8, GGTTATTTCCCTGAGCCA TKVDKKIEPRGPTIKPCP 3245- GTGACCTTGACCTGGAAC PCKCPAPNLLGGPSVFIF 3251 TCTGGATCCCTGTCGAGT PPKIKDVLMISLSPMVTC (1989). GGTGTGCACACCTTCCCA VVVDVSEDDPDVQISWF PMID: GCTGTCCTGCAGTCTGAC VNNVEVLTAQTQTHRED 2510996 CTCTACACCCTCAGCAGC YNSTLRVVSALPIQHQD TCAGTGACTGTAACCTCG WMSGKEFKCKVNNKAL AGCACCTGGCCCAGCCAG PAPIERTLSKPKGSVRAP TCCATCACCTGCAATGTG QVYVLPPPEEEMTKKQV GCCCACCCGGCAAGCAG TLTCMVTDFMPEDIYVE CACCAAGGTGGACAAGAA WTNNGKTELNYKNTEPV AATTGAGCCCAGAGGGCC LDSDGSYFMYSKLRVEK CACAATCAAACCCTGTGC KNWVERNSYSCSVVHEG TCCATGCAAATGCCCAGC LHNHHTTKSFSRITGK ACCTAACCTCTTGGGTGG (SEQ ID NO: 50) ACCATCCGTCTTCATCTT CCCTCCAAAGATCAAGGA TGTACTCATGATCTCCCT GAGTCCCATGGTCACATG TGTGGTGGTGGATGTGAG CGAGGATGACCCAGATGT CCAGATCAGCTGGTTCGT GAACAACGTGGAAGTACT CACAGCTCAGACACAAAC CCATAGAGAGGATTACAA CAGTACTCTCCGGGTGGT CAGTGCCCTCCCCATCCA GCACCAGGACTGGATGA GTGGCAAGGAGTTCAAAT GCAAGGTCAACAACAAAG CCCTCCCAGCGCCCATCG AGAGAACCATCTCAAAAC CCAAAGGGTCAGTAAGAG CTCCACAGGTATATGTCT TGCCTCCACCAGAAGAAG AGATGACTAAGAAACAGG TCACTCTGACCTGCATGG TCACAGACTTCATGCCTG AAGACATTTACGTGGAGT GGACCAACAACGGGAAA ACAGAGCTAAACTACAAG AACACTGAACCAGTCCTG GACTCTGATGGTTCTTAC TTCATGTACAGCAAGCTG AGAGTGGAAAAGAAGAA CTGGGTGGAAAGAAATAG CTACTCCTGTTCAGTGGT CCACGAGGGTCTGCACAA TCACCACACGACTAAGAG CTTCTCCCGGACTCCGGG TAAATGA (SEQ ID NO: 51) IgG2b GCCAAAACAACACCCCCA AKTTPPSVYPLAPGCGDT J00461 Yamawak vari- TCAGTCTATCCACTGGCC TGSSVTLGCLVKGYFPES AH00530 i- ant 1 CCTGGGTGTGGAGATACA VTVTWNSGSLSSSVHTFP 9  Kataoka  ACTGGTTCCTCCGTGACT ALLQSGLYTMSSSVTVPS V00801 Y. CTGGGATGCCTGGTCAAG STWPSQTVTCSVAHPASS D78344 et al., GGCTACTTCCCTGAGTCA TTVDKKLEPSGPISTINP Nature, GTGACTGTGACTTGGAAC CPPCKECHKCPAPNLEG 283, TCTGGATCCCTGTCCAGC GPSVFIFPPNIKDVLMIS 786-789 AGTGTGCACACCTTCCCA LTPKVTCVVVDVSEDDPD (1980). GCTCTCCTGCAGTCTGGA VQISWFVNNVEVHTAQT PMID: CTCTACACTATGAGCAGC QTHREDYNSTIRVVSTLP 6766534 TCAGTGACTGTCCCCTCC IQHQDWMSGKEFKCKV Ollo R. AGCACTTGGCCAAGTCAG NNKDLPSPIERTISKIKG and ACCGTCACCTGCAGCGTT LVRAPQVYILPPPAEQLS Rougeon GCTCACCCAGCCAGCAGC RKDVSLTCLVVGFNPGDI F., ACCACGGTGGACAAAAAA SVEWTSNGHTEENYKDTA Nature, CTTGAGCCCAGCGGGCCC PVLDSDGSYFIYSKLNMK 296, ATTTCAACAATCAACCCC TSKWEKTDSFSCNVRHE 761-763 TGTCCTCCATGCAAGGAG GLKNYYLKKTISRSPGK (1982). TGTCACAAATGCCCAGCT (SEQ ID NO: 52) PMID: CCTAACCTCGAGGGTGGA 6803173 CCATCCGTCTTCATCTTC Akahori CCTCCAAATATCAAGGAT Y. and GTACTCATGATCTCCCTG Kurosawa ACACCCAAGGTCACGTGT Y., GTGGTGGTGGATGTGAG Geno- CGAGGATGACCCAGACGT mics., CCAGATCAGCTGGTTTGT 41, GAACAACGTGGAAGTACA 100-104 CACAGCTCAGACACAAAC (1997). CCATAGAGAGGATTACAA PMID: CAGTACTATCCGGGTGGT 9126488 CAGCACCCTCCCCATCCA GCACCAGGACTGGATGA GTGGCAAGGAGTTCAAAT GCAAGGTCAACAACAAAG ACCTCCGATCACCGATCG AGAGAACCATCTCAAAAA TTAAAGGGCTAGTCAGAG CTCCACAAGTATACATCT TGCCGCCACCAGCAGAGC AGTTGTCCAGGAAAGATG TCAGTCTCACTTGCCTGG TCGTGGGCTTCAACCCTG GAGACATCAGTGTGGAGT GGACCAGCAATGGGCATA CAGAGGAGAACTACAAG GACACCGCACCAGTCCTA GACTCTGACGGTTCTTAC TTCATATATAGCAAGCTC AATATGAAAACAAGCAAG TGGGAGAAAACAGATTCC TTCTCATGCAACGTGAGA CACGAGGGTCTGAAAAAT TACTACCTGAAGAAGACC ATCTCCCGGTCTCCGGGT AAATGA (SEQ ID NO: 53) IgG2b GCCAAAACAACACCCCCA AKTTPPSVYPLAPGCGDT V00763 Tucker vari- TCAGTCTATCCACTGGCC TGSSVTSGCLVKGYFPEP P. W. et ant 2 CCTGGGTGTGGAGATACA VTVTWNSGSLSSSVHTFP al., ACTGGTTCCTCCGTGACC ALLQSGLYTMSSSVTVPS Sci- TCTGGGTGCCTGGTCAAG STWPSQTVTCSVAHPASS ence., GGGTACTTCCCTGAGCCA TTVDKKLEPSGPISTINP 206, GTGACTGTGACTTGGAAC CPPCKECHKCPAPNLEG 1303- TCTGGATCCCTGTCCAGC GPSVFIFPPNIKDVLMIS 1306 AGTGTGCACACCTTCCCA LTPKVTCVVVDVSEDDPD (1979). GCTCTCCTGCAGTCTGGA VQISWFVNNVEVHTAQT PMID: CTCTACACTATGAGCAGC QTHREDYNSTIRVVSTLP 117549 TCAGTGACTGTCCCCTCC IQHQDWMSGKEFKCKV AGCACCTGGCCAAGTCAG NNKDLPSPIERTISKIKG ACCGTCACCTGCAGCGTT LVRAPQVYTLPPPAEQLS GCTCACCCAGCCAGCAGC RKDVSLTCLVVGFNPGDI ACCACGGTGGACAAAAAA SVEWTSNGHTEENYKDTA CTTGAGCCCAGCGGGCCC PVLDSDGSYFIYSKLNMK ATTTCAACAATCAACCCC TSKWEKTDSFSCNVRHE TGTCCTCCATGCAAGGAG GLKNYYLKKTISRSPGK TGTCACAAATGCCCAGCT (SEQ ID NO: 54) CCTAACCTCGAGGGTGGA CCATCCGTCTTCATCTTC CCTCCAAATATCAAGGAT GTACTCATGATCTCCCTG ACACCCAAGGTCACGTGT GTGGTGGTGGATGTGAG CGAGGATGACCCAGACGT CCAGATCAGCTGGTTTGT GAACAACGTGGAAGTACA CACAGCTCAGACACAAAC CCATAGAGAGGATTACAA CAGTACTATCCGGGTGGT CAGCACCCTCCCCATCCA GCACCAGGACTGGATGA GTGGCAAGGAGTTCAAAT GCAAGGTGAACAACAAAG ACCTCCCATCACCCATCG AGAGAACCATCTCAAAAA TTAAAGGGCTAGTCAGAG CTCCACAAGTATACACTT TGCCGCCACCAGCAGAGC AGTTGTCCAGGAAAGATG TCAGTCTCACTTGCCTGG TCGTGGGCTTCAACCCTG GAGACATCAGTGTGGAGT GGACCAGCAATGGGCATA CAGAGGAGAACTACAAG GACACCGCACCAGTTCTT GACTCTGACGGTTCTTAC TTCATATATAGCAAGCTC AATATGAAAACAAGCAAG TGGGAGAAAACAGATTCC TTCTCATGCAACGTGAGA CACGAGGGTCTGAAAAAT TACTACCTGAAGAAGACC ATCTCCCGGTCTCCGGGT AAATGA (SEQ ID NO: 55) IgG2c GCCAAAACAACAGCCCCA AKTTAPSVYPLAPVCGGT J00479 Ollo R. vari- TCGGTCTATCCACTGGCC TGSSVTLGCLVKGYFPEP and ant 1 CCTGTGTGTGGAGGTACA VTLTWNSGSLSSGVHTFP Rougeon ACTGGCTCCTCGGTGACT ALLQSGLYTLSSSVTVTS F.,  CTAGGATGCCTGGTCAAG NTWPSQTITCNVAHPASS Cell, GGTTATTTCCCTGAGCCA TKVDKKIEPRVPITQNPC 32, GTGACCTTGACCTGGAAC PPLKECPPCAAPDLLGGP 515-523 TCTGGATCCCTGTCCAGT SVFIFPPKIKDVLMISLS (1983). GGTGTGCACACCTTCCCA PMVTCVVVDVSEDDPDVQ PMID: GCTCTCCTGCAGTCTGGC ISWFVNNVEVHTAQTQT 6297797 CTCTACACCCTCAGCAGC HREDYNSTLRVVSALPIQ TCAGTGACTGTAACCTCG HQDWMSGKEFKCKVNN AACACCTGGCCCAGCCAG RALPSPIEKTISKPRGPV ACCATCACCTGCAATGTG RAPQVYVLPPPAEEMTKK GCCCACCCGGCAAGCAG EFSLTCMITGFLPAEIAV CACCAAAGTGGACAAGAA DWTSNGRTEQNYKNTAT AATTGAGCCCAGAGTGCC VLDSDGSYFMYSKLRVQ CATAACACAGAACCCCTG KSTWERGSLFACSVVHE TCCTCCACTCAAAGAGTG VLHNHLTTKTISRSLGK TCCCCCATGCGCAGCTCC (SEQ ID NO: 56) AGACCTCTTGGGTGGACC ATCCGTCTTCATCTTCCC TCCAAAGATCAAGGATGT ACTCATGATCTCCCTGAG CCCCATGGTCACATGTGT GGTGGTGGATGTGAGCG AGGATGACCCAGACGTCC AGATCAGCTGGTTTGTGA ACAACGTGGAAGTACACA CAGCTCAGACACAAACCC ATAGAGAGGATTACAACA GTACTCTCCGGGTGGTCA GTGCCCTCCCCATCCAGC ACCAGGACTGGATGAGTG GCAAGGAGTTCAAATGCA AGGTCAACAACAGAGCCC TCCCATCCCCCATCGAGA AAACCATCTCAAAACCCA GAGGGCCAGTAAGAGCT CCACAGGTATATGTCTTG CCTCCACCAGCAGAAGAG ATGACTAAGAAAGAGTTC AGTCTGACCTGCATGATC ACAGGCTTCTTACCTGCC GAAATTGCTGTGGACTGG ACCAGCAATGGGCGTACA GAGCAAAACTACAAGAAC ACCGCAACAGTCCTGGAC TCTGATGGTTCTTACTTC ATGTACAGCAAGCTCAGA GTACAAAAGAGCACTTGG GAAAGAGGAAGTCTTTTC GCCTGCTCAGTGGTCCAC GAGGTGCTGCACAATCAC CTTACGACTAAGACCATC TCCCGGTCTCTGGGTAAA TGA (SEQ ID NO: 57) IgG2c GCCAAAAGAACAGCCCCA AKTTAPSVYPLAPVCGGT X16998 Morgado vari- TCGGTCTATCCACTGGCC TGSSVTLGCLVKGYFPEP M. G. et ant 2 CCTGTGTGTGGAGGTACA VTLTWNSGSLSSGVHTFP al., ACTGGCTCCTCGGTGACT ALLQSGLYTLSSSVTVTS EMBO J., CTAGGATGCCTGGTCAAG NTWPSQTITCNVAHPASS 8, GGTTATTTCCCTGAGCCA TKVDKKIESRRPIPPNSC 3245- GTGACCTTGACCTGGAAC PPCKECSIFPAPDLLGGP 3251 TCTGGATCCCTGTCCAGT SVFIFPPKIKDVLMISLS (1989). GGTGTGCACACCTTCCCA PIVTCVVVDVSEDDPDVQ PMID: GCTCTCCTGCAGTCTGGC ISWFVNNVEVHTAQTQTH 2510996 CTCTACACCCTCAGCAGC REDYNSTLRVVSALPIQH TCAGTGACTGTAACCTCG QDWMSGKEFKCKVNNR AACACCTGGCCCAGCCAG ALPSPIEKTISKPRGPVR ACCATCACCTGCAATGTG APQVYVLPPPAEEMTKKE GCCCACCCGGCAAGCAG FSLTCMITDFLPAEIAVD CACCAAAGTGGACAAGAA WTSNGHKELNYKNTAPV AATTGAATCCAGAAGGCC LDTDGSYFMYSKLRVQK CATACCACCCAACTCCTG STWEKGSLFACSVVHEG TCCTCCATGCAAAGAGTG LHNHHTTKTISRSLGK TTCCATATTCCCAGCTCC (SEQ ID NO: 58) TGACCTCTTGGGTGGACC ATCCGTCTTCATCTTCCC TCCAAAGATCAAGGATGT ACTCATGATCTCCCTGAG CCCCATAGTCACATGTGT GGTGGTGGATGTGAGCG AGGATGACCCAGATGTCC AGATCAGCTGGTTTGTGA ACAACGTGGAAGTACACA CAGCTCAGACACAAACCC ATAGAGAGGATTACAACA GTACTCTCCGGGTGGTCA GTGCCCTCCCCATCCAGC ACCAGGACTGGATGAGTG GCAAGGAGTTCAAATGCA AGGTCAACAACAGAGCCC TCCCATCCCCCATCGAGA AAACCATCTCAAAACCCA GAGGGCCAGTAAGAGCT CCACAGGTATATGTCTTG CCTCCACCAGCAGAAGAG ATGACTAAGAAAGAGTTC AGTCTGACCTGCATGATC ACAGACTTCTTACCTGCC GAAATTGCTGTGGACTGG ACCAGCAATGGGCATAAA GAGCTGAACTACAAGAAC ACCGCACCAGTCCTGGAC ACTGATGGTTCTTACTTC ATGTACAGCAAGCTCAGA GTGCAAAAGAGCACTTGG GAAAAAGGAAGTCTTTTC GCCTGCTCAGTGGTCCAC GAGGGTCTGCACAATCAC CATACGACTAAGACCATC TCCCGGTCTCTGGGTAAA TGA (SEQ ID NO: 59) IgG2c GCCAAAACAACAGCCCCA AKTTAPSVYPLAPVCGGT Y10606 Martin vari- TCGGTCTATCCACTGGCC TGSSVTLGCLVKGYFPEP R. M. et ant 3 CCTGTGTGTGGAGGTACA VTLTWNSGSLSSGVHTFP al., ACTGGCTCCTCGGTGACT ALLQSGLYTLSSSVTVTS Immuno- CTAGGATGCCTGGTCAAG NTWPSQTITCNVAHPASS gene- GGTTATTTCCCTGAGCCA TKVDKKIEPRVPITQNPC tics, GTGACCTTGACCTGGAAC PPLKECPPCAAPDLLGGP 46, TCTGGATCCCTGTCCAGT SVFIFPPKIKDVLMISLS 167-168 GGTGTGCACACCTTCCCA PMVTCVVVDVSEDDPDVQ (1997). GCTCTCCTGCAGTCTGGC ISWFVNNVEVHTAQTQT PMID: CTCTACACCCTCAGCAGC HREDYNSTLRVVSALPIQ 9162106 TCAGTGACTGTAACCTCG HQDWMSGKEFKCKVNN AACACCTGGCCCAGCCAG RALPSPIEKTISKPRGPV ACCATCACCTGCAATGTG RAPQVYVLPPPAEEMTKK GCCCACCCGGCAAGCAG EFSLTCMITGFLPAEIAV CACCAAAGTGGACAAGAA DWTSNGRTEQNYKNTAT AATTGAGCCCAGAGTGCC VLDSDGSYFMYSKLRVQ CATAACACAGAACCCCTG KSTWERGSLFACSVVHE TCCTCCACTCAAAGAGTG GLHNHLTTKTISRSLGK TCCCCCATGCGCAGCTCC (SEQ ID NO: 60) AGACCTCTTGGGTGGACC ATCCGTCTTCATCTTCCC TCCAAAGATCAAGGATGT ACTCATGATCTCCCTGAG CCCCATGGTCACATGTGT GGTGGTGGATGTGAGCG AGGATGACCCAGACGTCC AGATCAGCTGGTTTGTGA ACAACGTGGAAGTACACA CAGCTCAGACACAAACCC ATAGAGAGGATTACAACA GTACTCTCCGGGTGGTCA GTGCCCTCCCCATCCAGC ACCAGGACTGGATGAGTG GCAAGGAGTTCAAATGCA AGGTGAACAACAGAGCCC TCCCATCCCCCATCGAGA AAACCATCTCAAAACCCA GAGGGCCAGTAAGAGCT CCACAGGTATATGTCTTG CCTCCACCAGCAGAAGAG ATGACTAAGAAAGAGTTC AGTCTGACCTGCATGATC ACAGGCTTCTTACCTGCC GAAATTGCTGTGGACTGG ACCAGCAATGGGCGTACA GAGCAAAACTACAAGAAC ACCGCAACAGTCCTGGAC TCTGATGGTTCTTACTTC ATGTACAGCAAGCTCAGA GTACAAAAGAGCACTTGG GAAAGAGGAAGTCTTTTC GCCTGCTCAGTGGTCCAC GAGGGTCTGCACAATCAC CTTACGACTAAGACCATC TCCCGGTCTCTGGGTAAA TGA (SEQ ID NO: 61) IgG3 GCTACAACAACAGCCCCA ATTTAPSVYPLVPGCSDT J00451 Stanton TCTGTCTATCCCTTGGTC SGSSVTLGCLVKGYFPEP AH00530 L. W.  CCTGGCTGCAGTGACACA VTVKWNYGALSSGVRTV 9  and TCTGGATCCTCGGTGACA SSVLQSGFYSLSSLVTVP X00915 Marcu CTGGGATGCCTTGTCAAA SSTWPSQTVICNVAHPAS D78343 K. B., GGCTACTTCCCTGAGCCG KTELIKRIEPRIPKPSTP Nucleic GTAACTGTAAAATGGAAC PGSSCPPGNILGGPSVFI Acids  TATGGAGCCCTGTCCAGC FPPKPKDALMISLTPKVT Res., GGTGTGCGCACAGTCTCA CVVVDVSEDDPDVHVSWF 10, TCTGTCCTGCAGTCTGGG VDNKEVHTAWTQPREAQY 5993- TTCTATTCCCTCAGCAGC NSTFRVVSALPIQHQDW 6006 TTGGTGACTGTACCCTCC MRGKEFKCKVNNKALPA (1982). AGCACCTGGCCCAGCCAG PIERTISKPKGRAQTPQV PMID: ACTGTCATCTGCAACGTA YTIPPPREQMSKKKVSLT 6292864 GCCCACCGAGCCAGCAAG CLVTNFFSEAISVEWERN Wels ACTGAGTTGATCAAGAGA GELEQDYKNTPPILDSDG J. A. ATCGAGCCTAGAATACCC TYFLYSKLTVDTDSWLQ et al., AAGCCCAGTACCCCCCCA GEIFTCSVVHEALHNHH EMBO J., GGTTCTTCATGCCCACCT TQKNLSRSPGK 3, GGTAACATCTTGGGTGGA (SEQ ID NO: 62) 2041- CCATCCGTCTTCATCTTC 2046 CCCCCAAAGCCCAAGGAT (1984). GCACTCATGATCTCCCTA PMID: ACCCCCAAGGTTACGTGT 6092053 GTGGTGGTGGATGTGAG Akahori CGAGGATGACCCAGATGT Y. and CCATGTCAGCTGGTTTGT Kurosawa GGACAACAAAGAAGTACA Y., CACAGCCTGGACACAGCC Geno- CCGTGAAGCTCAGTACAA mics., CAGTACCTTCCGAGTGGT 41, CAGTGCCCTCCCCATCCA 100-104 GCACCAGGACTGGATGA (1997). GGGGCAAGGAGTTCAAAT PMID: GCAAGGTCAACAACAAAG 9126488 CCCTCCCAGCCCCCATCG AGAGAACCATCTCAAAAC CCAAAGGAAGAGCCCAG ACACCTCAAGTATACACC ATACCCCCACCTCGTGAA CAAATGTCCAAGAAGAAG GTTAGTCTGACCTGCCTG GTCACCAACTTCTTCTCT GAAGCCATCAGTGTGGAG TGGGAAAGGAACGGAGA ACTGGAGCAGGATTACAA GAACACTCCACCCATCCT GGACTCAGATGGGACCTA CTTCCTCTAGAGCAAGCT CACTGTGGATACAGACAG TTGGTTGCAAGGAGAAAT TTTTACCTGCTCCGTGGT GCATGAGGCTCTCCATAA CCACCACACACAGAAGAA CCTGTCTCGCTCCCCTGG TAAATGA (SEQ ID NO: 63) Mouse Ig GCTGATGCTGCACCAACT ADAAPTVSIFPPSSEQLT V00807 http:// Hieter Ig kappa GTATCCATCTTCCCACCA SGGASVVCFLNNFYPKDI V00777 www. P. A.  light (CK) TCCAGTGAGCAGTTAACA NVKWKIDGSERQNGVLN V01569 imgt.org/ et al.,  chain TCTGGAGGTGCCTCAGTC SWTDQDSKDSTYSMSST V00806 IMGT Cell,  con- GTGTGCTTCTTGAACAAC LTLTKDEYERHNSYTCE X67002 reper 22, stant TTCTACCCCAAAGACATC ATHKTSTSPIVKSFNRN X67003 toire/ 197-207 re- AATGTCAAGTGGAAGATT EC  X67004 index.php? (1980). gion GATGGCAGTGAACGACAA (SEQ ID NO: 36) X67005 section = PMID: AATGGCGTCCTGAACAGT X67006 Locus 6775818  TGGACTGATCAGGACAGC X67007 Genes & Max  AAAGACAGCACCTACAGC X67008 reper E. E. ATGAGCAGCACCCTCACG X67009 toire = et al., TTGACCAAGGACGAGTAT X67010 gene J. Biol. GAACGACATAACAGCTAT X67011 table & Chem., ACCTGTGAGGCCACTCAC X67012 species = 256, AAGACATCAACTTCACCC Mus_ 5116- ATTGTCAAGAGCTTCAAC musculus & 5120 AGGAATGAGTGTTAG group = (1981). (SEQ ID NO: 37) IGKC PMID: 6262318 Seidman J. G., et al., Nature, 280, 370-375 (1979). PMID: 111146 Solin M. L. and Kaarti- nen Immuno- gene- tics, 37, 401-407 (1993). PMID: 8436414 Ig GGCCAGCCCAAGTCTTCG GQPKSSPSVTLFPPSSEE J00587 http:// Selsing  lambda CCATCAGTCACCCTGTTT LETNKATLVCTITDFYPG AH00531 www. E. 1 (CL) CCACCTTCCTCTGAAGAG VVTVDWKVDGTPVTQG 1  imgt.org/ et al., CTCGAGACTAACAAGGCC METTQPSKQSNNKYMAS X58411 IMGT Proc. ACACTGGTGTGTACGATC SYLTLTARAWERHSSYSC V00814 reper Natl.  ACTGATTTCTACCCAGGT QVTHEGHTVEKSLSRAD toire/ Acad. GTGGTGACAGTGGACTG CS (SEQ ID NO: 38) index.php? Sci. GAAGGTAGATGGTACCCC section = USA, 79, TGTCACTCAGGGTATGGA Locus 4681- GACAACCCAGCCTTCCAA Genes & 4685 ACAGAGCAACAACAAGTA reper (1982). CATGGCTAGCAGCTACCT toire = PMID: GACCCTGACAGCAAGAGC gene 6812053 ATGGGAAAGGCATAGCA table & Weiss S. GTTACAGCTGCCAGGTCA species = and Wu CTCATGAAGGTCACACTG Mus_ GE, TGGAGAAGAGTTTGTCCC musculus & EMBO J., GTGCTGACTGTTCCTAG group = 6, (SEQ ID NO: 39) IGLC 927-937 (1987). PMID: 3109891 Bernard O. et al., Cell,  15, 1133- 1144 (1978). PMID: 103630 Ig GGTCAGCCCAAGTCCACT GQPKSTPTLTVFPPSSEE J00595 Selsing  lambda CCCACTCTCACCGTGTTT LKENKATLVCLISNFSPS AH00196 E. 2 (CL) CCACCTTCCTCTGAGGAG GVTVAWKANGTPITQGV 8  et al., CTCAAGGAAAACAAAGCC DTSNPTKEGNKFMASSF J00592 Proc. ACACTGGTGTGTCTGATT LHLTSDQWRSHNSFTCQ AH00196  Natl.  TCCAACTTTTCCCCGAGT VTHEGDTVEKSLSPAECL 7 Acad. GGTGTGACAGTGGCCTG (SEQ ID NO: 40) X58414 Sci. GAAGGCAAATGGTACACC USA, 79, TATCACCCAGGGTGTGGA 4681- CACTTCAAATCCCACCAA 4685 AGAGGGCAACAAGTTCAT (1982).  GGCCAGCAGCTTCCTACA PMID:  TTTGACATCGGACCAGTG 6812053  GAGATCTCACAACAGTTT Wu G. TACCTGTCAAGTTACACA et al., TGAAGGGGACACTGTGG Cell, AGAAGAGTCTGTCTCCTG 33, CAGAATGTCTCTAA 77-83 (SEQ ID NO: 41) (1983). PMID: 6432336 Weiss S. and Wu GE, EMBO J., 6, 927-937 (1987). PMID: 3109891 Ig GGTCAGCCCAAGTCCACT GQPKSTPTLTMFPPSPEE J00585 Selsing  lambda CCCACACTCACCATGTTT LQENKATLVCLISNFSPS AH00531 E. 3 (CL) CCACCTTCCCCTGAGGAG GVTVAWKANGTPITQGV 1  et al., CTCCAGGAAAACAAAGCC DTSNPTREDNKYMASSF X58415 Proc. ACACTCGTGTGTCTGATT LHLTSDQWRSHNSFTCQ X58411 Natl.  TCCAATTTTTCCCCAAGT VTHEGDTVEKSLSPAECL Acad. GGTGTGACAGTGGCCTG (SEQ ID NO: 42) Sci. GAAGGCAAATGGTACACC USA, 79, TATCACCCAGGGTGTGGA 4681- CACTTCAAATCCCACCAA 4685 AGAGGACAACAAGTACAT (1982). GGCCAGCAGCTTCTTACA PMID: TTTGACATCGGACCAGTG 6812053 GAGATCTCACAACAGTTT Weiss S. TACCTGCCAAGTTACACA and Wu TGAAGGGGACACTGTGG GE, AGAAGAGTCTGTCTCCTG EMBO J., CAGAATGTCTCTAA 6, (SEQ ID NO: 43) 927-937 (1987). PMID: 3109891 Bovine Bovine IgG1 GCCTCCACCACAGCCCCG ASTTAPKVYPLSSCCGDKSSST X62916 http:// Symons (Scien- Ig vari- AAAGTCTACCCTCTGAGTT VTLGCLVSSYMPEPVTVTWNS www. D. B.  tific heavy ant 1 CTTGCTGCGGGGACAAGT GALKSGVHTFPAVLQSSGLYSL imgt.org/ et al.,  Name: chain CCAGCTCCACCGTGACCC SSMVTVPGSTSGQTFTCNVAHP IMGT J. Bos con- TGGGCTGCCTGGTCTCCA ASSTKVDKAVDPTCKPSPCDCC reper Immuno-  taurus) stant GCTACATGCCCGAGCCGG PPPELPGGPSVFIFPPKPKDTLT toire/ genet., region TGACCGTGACCTGGAACT ISGTPEVTCVVVDVGHDDPEVK index.php? 14, (CH1~ CGGGTGCCCTGAAGAGCG FSWFVDDVEVNTATTKPREEQ section = 273-283 CH3) GCGTGCACACCTTCCCGG FNSTYRVVSALRIQHQDWTGG Locus (1987). CTGTCCTTCAGTCCTCCGG KEFKCKVHNEGLPAPIVRTISR Genes & PMID: GCTGTACTCTCTCAGCAG TKGPAREPQVYYLAPPQEELSK reper 3141517  CATGGTGACCGTGCCCGG STVSLTCMVTSFYPDYIAVEWQ toire = Symons  CAGCACCTCAGGACAGAC RNGQPESEDKYGTTPPQLDAD gene D. B. CTTCACCTGCAACGTAGC SSYFLYSKLRVDRNSWQEGDT table & et al., CCACCCGGCCAGCAGCAC YTCVVMHEALHNHYTQKSTSK species = Mol. CAAGGTGGACAAGGCTGT SAGE* (SEQ ID NO: 66) bovine & Immun- TGATCCCACATGCAAACC group = ol., ATCACCCTGTGACTGTTGC IGHC 26, CCACCCCCTGAGCTCCCC 841-850 GGAGGACCCTCTGTCTTC (1989). ATCTTCCCACGAAACCCA PMID: AGGACACCCTCACAATCT 2513487  CGGGAACGCCCGAGGTCA Kacsko- CGTGTGTGGTGGTGGACG vics TGGGCCACGATGACCCCG I. and AGGTGAAGTTCTCCTGGT Butler TCGTGGACGACGTGGAGG J. E., TAAACACAGCCACGACGA Mol. AGCCGAGAGAGGAGCAGT Immun- TCAACAGCACCTACCGCG ol.,  TGGTCAGCGCCCTGCGCA 33, TCCAGCACCAGGACTGGA 189-195 CTGGAGGAAAGGAGTTCA (1996). AGTGCAAGGTCCACAACG PMID: AAGGCCTCCCGGCCCCCA 8649440 TCGTGAGGACCATCTCCA Rabbani GGACCAAAGGGCCGGCCC H. GGGAGCCGCAGGTGTATG et al.,  TCCTGGCCCCACCCCAGG Immuno- AAGAGCTCAGCAAAAGCA gene- CGGTCAGCCTCACCTGCA tics, TGGTCACCAGCTTCTACCC 46, AGACTACATCGCCGTGGA 326-331 GTGGCAGAGAAACGGGCA (1997). GCCTGAGTCGGAGGACAA PMID: GTACGGCACGACCCCGCC 9218535 CCAGCTGGACGCCGACAG Saini CTCCTACTTCCTGTAGAGC S. S. AAGCTCAGGGTGGACAGG et al., AACAGCTGGCAGGAAGGA Scand.  GACACCTACACGTGTGTG J. GTGATGCACGAGGCCCTG Immunol. CACAATCACTACACGCAG 65, 32-8 AAGTCCACCTCTAAGTCTG (2007). CGGGTAAATGA PMID: (SEQ ID NO: 67) 17212764 IgG1 GCCTCGACCACAGCCCCG ASTTAPKVYPLSSCCGDKSSST X16701 vari- AAAGTCTACCCTCTGAGTT VTLGCLVSSYMPEPVTVTWNS (M25278) ant 2 CTTGCTGCGGGGACAAGT GALKSGVHTFPAVLQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPGSTSGQTFTCNVAHP TGGGCTGCCTGGTCTCCA ASSTKVDKAVDPTCKPSPCDCC GCTACATGCCCGAGCCGG PPPELPGGPSVFIFPPKPKDTLT TGACCGTGACCTGGAACT ISGTPEVTCVVVDVGHDDPEVK CGGGTGCCCTGAAGAGCG FSWFVDDVEVNTATTKPREEQ GCGTGCACACCTTCCCGG FNSTYRVVSALRIQHQDWTGG CCGTCCTTCAGTCCTCCG KEFKCKVHNEGLPAPIVRTISR GGCTGTACTCTCTCAGCA TKGPAREPQVYVLAPPQEELSK GCATGGTGACCGTGCCCG STVSLTCMVTSFYPDYIAVEWQ GCAGCACCTCAGGACAGA RNGQPESEDKYGTTPPQLDAD CCTTCACCTGCAACGTAG SSYFLYSKLRVDRNSWQEGDT CCCACCCGGCCAGCAGCA YTCVVMHEALHNHYTQKSTSK CCAAGGTGGACAAGGCTG SAGK* (SEQ ID NO: 68) TTGATCCCACATGCAAACC ATCACCCTGTGACTGTTGC CCACCCCCTGAGCTCCCC GGAGGACCCTCTGTCTTC ATCTTCCCACCGAAACCCA AGGACACCCTCACAATCT CGGGAACGCCCGAGGTCA CGTGTGTGGTGGTGGACG TGGGCCACGATGACCCCG AGGTGAAGTTCTCCTGGT TCGTGGACGACGTGGAGG TAAACACAGCCACGACGA AGCCGAGAGAGGAGCAGT TCAACAGCACCTACCGCG TGGTCAGCGCCCTGCGCA TCCAGCACCAGGACTGGA CTGGAGGAAAGGAGTTCA AGTGCAAGGTCCACAACG AAGGCCTCCCGGCCCCCA TCGTGAGGACCATCTCCA GGACCAAAGGGCCGGCCC GGGAGCCGCAGGTGTATG TCCTGGCCCCACCCCAGG AAGAGCTCAGCAAAAGCA CGGTCAGCCTCACCTGCA TGGTCACCAGCTTCTACCC AGACTACATCGCCGTGGA GTGGCAGAGAAACGGGCA GCCTGAGTCGGAGGACAA GTACGGCACGACCCCGCC CCAGCTGGACGCCGACAG CTCCTACTTCCTGTACAGC AAGCTCAGGGTGGACAGG AACAGCTGGCAGGAAGGA GACACCTACACGTGTGTG GTGATGCACGAGGCCCTG CACAATCACTACACGCAG AAGTCCACCTCTAAGTCTG CGGGTAAATGA (SEQ ID NO: 69) IgG1 GCCTCCACCACAGCCCCG ASTTAPKVYPLSSCCGDKSSST S82409 vari- AAAGTCTACCCTCTGAGTT VTLGCLVSSYMPEPVTVTWNS ant 3 CTTGCTGCGGGGACAAGT GALKSGVHTFPAVLQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPGSTSGTQTFTCNVAH TGGGCTGCCTGGTCTCCA PASSTKYDKAVDPRCKTTCDCC GCTACATGCCCGAGCCGG PPPELPGGPSVFIFPPKPKDTLT TGACCGTGACCTGGAACT ISGTPEVTCVVVDVGHDDPEVK CGGGTGCCCTGAAGAGCG FSWFVDDVEVNTATTKPREEQ GCGTGCACACCTTCCCGG FNSTYRVVSALRIQHQDWTGG CCGTCCTTCAGTCCTCCG KEFKCKVHNEGLPAPIVRTISR GGCTCTACTCTCTCAGCA TKGPAREPQVYVLAPPQEELSK GCATGGTGACCGTGCCCG STVSLTCMVTSFYPDYIAVEWQ GCAGCACCTCAGGAACCC RNGQPESEDKYGTTPPQLDAD AGACCTTCACCTGCAACG GSYFLYSRLRVDRNSWQEGDT TAGCCCACCCGGCCAGCA YTCVVMHEALHNHYTQKSTSK GCACCAAGGTGGACAAGG SAGK* CTGTTGATCCCAGATGCA (SEQ ID NO: 70) AAACAACCTGTGACTGTT GCCCACCGCCTGAGCTCC CTGGAGGACCCTCTGTCT TCATCTTCCCACCGAAACC CAAGGACACCCTCACAAT CTCGGGAACGCCCGAGGT CACGTGTGTGGTGGTGGA CGTGGGCCACGATGACCC CGAGGTGAAGTTCTCCTG GTTCGTGGACGACGTGGA GGTAAACACAGCCACGAC GAAGCCGAGAGAGGAGCA GTTCAACAGCACCTACCG CGTGGTCAGCGCCCTGCG CATCCAGCACCAGGACTG GACTGGAGGAAAGGAGTT CAAGTGCAAGGTCCACAA CGAAGGCCTCCCAGCCCC CATCGTGAGGACCATCTC CAGGACCAAAGGGCCGGC CCGGGAGCCGCAGGTGTA TGTCCTGGCCCCACCCCA GGAAGAGCTCAGCAAAAG CACGGTCAGCCTCACCTG CATGGTCACCAGCTTCTAC CCAGACTACATCGCCGTG GAGTGGCAGAGAAATGGG CAGCCTGAGTCAGAGGAC AAGTACGGCACGACCCCT CCCCAGCTGGACGCCGAC GGCTCCTACTTCCTGTACA GCAGGCTCAGGGTGGACA GGAACAGCTGGCAGGAAG GAGACACCTACACGTGTG TGGTGATGCACGAGGCCC TGCACAATCACTACACGC AGAAGTCCACCTCTAAGT CTGCGGGTAAATGA (SEQ ID NO: 71) IgG2 GCCTCCACCACAGCCCCG ASTTAPKVYPLASSCGDTSSST S82407 vari- AAAGTCTACCCTCTGGCAT VTLGCLVSSYMPEPVTVTWNS ant 1 CCAGCTGCGGAGACACAT GALKSGVHTFPAVLQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPASSSGQTFTCNVAHP TGGGCTGCCTGGTGTCCA ASSTKVDKAVGVSIDCSKCHNQ GCTACATGCCCGAGCCGG PCVREPSVFIFPPKPKDTLMITG TGACCGTGACCTGGAACT TPEVTCVVVNVGHDNPEVQFS CGGGTGCCCTGAAGAGCG WFVDDVEVHTARSKPREEQFN GCGTGCACACCTTCCCGG STYRVVSALPIQHQDWTGGKE CTGTCCTTCAGTCCTCCGG FKCKVNNKGLSAPIVRIISRSKG GCTCTACTCTCTCAGCAGC PAREPQVYVLDPPKEELSKSTL ATGGTGACCGTGCCCGCC SVTCMVTGFYPEDVAVEWQRN AGCAGCTCAGGACAGACC RQTESEDKYRTTPPQLDTDRSY TTCACCTGCAACGTAGCC FLYSKLRVDRNSWQEGDAYTC CACCCGGCCAGCAGCACC VVMHEALHNHYMQKSTSKSA AAGGTGGACAAGGCTGTT GK* (SEQ ID NO: 72) GGGGTCTCCATTGACTGC TCCAAGTGTCATAACCAG CCTTGCGTGAGGGAACCA TCTGTCTTCATCTTCCCAC CGAAACCCAAAGACACCC TGATGATCACAGGAACGC CCGAGGTCACGTGTGTGG TGGTGAACGTGGGCCACG ATAACCCCGAGGTGCAGT TCTCCTGGTTCGTGGATG ACGTGGAGGTGCACACGG CCAGGTCGAAGCCAAGAG AGGAGCAGTTCAACAGCA CGTACCGCGTGGTCAGCG CCCTGCCCATCCAGCACC AGGACTGGACTGGAGGAA AGGAGTTCAAGTGCAAGG TCAACAACAAAGGCCTCT CGGCCCCCATCGTGAGGA TCATCTCCAGGAGCAAAG GGCCGGCCCGGGAGCCGC AGGTGTATGTCCTGGACC CACCCAAGGAAGAGCTCA GCAAAAGCACGCTCAGCG TCACCTGCATGGTCACCG GCTTCTACCCAGAAGATG TAGCCGTGGAGTGGCAGA GAAACCGGCAGACTGAGT CGGAGGACAAGTACCGCA CGACCCCGCCCCAGCTGG ACACCGACCGCTCCTACTT CCTGTACAGCAAGCTCAG GGTGGACAGGAACAGCTG GCAGGAAGGAGACGCCTA CACGTGTGTGGTGATGCA CGAGGCCCTGCACAATCA CTACATGCAGAAGTCCAC CTCTAAGTCTGCGGGTAA ATGA (SEQ ID NO: 73) IgG2 GCCTCCACCAGAGCCCCG ASTTAPKVYPLSSCCGDKSSST M36946 vari- AAAGTCTACCCTCTGAGTT VTLGCLVSSYMPEPVTVTWNS (X06703) ant 2 CTTGCTGCGGGGACAAGT GALKSGVHTFPAVLQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPGSTSGQTFTCNVAHP TGGGCTGCCTGGTGTCCA ASSTKVDKAVGVSSDCSKPNN GCTACATGCCCGAGCCGG QHCVREPSVFIFPPKPKDTLMI TGACCGTGACCTGGAACT TGTPEVTCVVVNVGHDNPEVQ CGGGTGCCCTGAAGAGCG FSWFVDDVEVHTARTKPREEQ GCGTGCACACCTTCCCGG FNSTYRVVSALPIQHQDWTGG CCGTCCTTCAGTCCTCCG KEFKCKVNIKGLSASIVRIISRS GGCTCTACTCTCTCAGCA KGPAREPQVYVLDPPKEELSKS GCATGGTGACCGTGCCCG TVSVTCMVIGFYPEDVDVEWQ GCAGCACCTCAGGACAGA RDRQTESEDKYRTTPPQLDAD CCTTCACCTGCAACGTAG RSYFLYSKLRVDRNSWQRGDT CCCACCCGGCCAGCAGCA YTCVVMHEALHNHYMQKSTS CCAAGGTGGACAAGGCTG KSAGK* TTGGGGTCTCCAGTGACT (SEQ ID NO: 74) GCTCCAAGCCTAATAACC AGCATTGCGTGAGGGAAC CATCTGTCTTCATCTTCCC ACCGAAACCCAAAGACAC CCTGATGATCACAGGAAC GCCCGAGGTCACGTGTGT GGTGGTGAACGTGGGCCA CGATAACCCCGAGGTGCA GTTCTCCTGGTTCGTGGA CGACGTGGAGGTGCACAC GGCCAGGACGAAGCCGAG AGAGGAGCAGTTCAACAG CACGTACCGCGTGGTCAG CGCCCTGCCCATCCAGCA CCAGGACTGGACTGGAGG AAAGGAGTTCAAGTGCAA GGTCAACATCAAAGGCCT CTCGGCCTCCATCGTGAG GATCATCTCCAGGAGCAA AGGGCCGGCCCGGGAGCC GCAGGTGTATGTCCTGGA CCCACCCAAGGAAGAGCT CAGCAAAAGCACGGTCAG CGTGAGCTGCATGGTCAT CGGCTTCTACCCAGAAGA TGTAGACGTGGAGTGGCA GAGAGACCGGCAGACTGA GTCGGAGGACAAGTACCG CACGACCCCGCCCCAGCT GGACGCCGACCGCTCCTA CTTCCTGTACAGCAAGCTC AGGGTGGACAGGAACAGC TGGCAGAGAGGAGACACC TACACGTGTGTGGTGATG CACGAGGCCCTGCACAAT CACTACATGCAGAAGTCC ACCTCTAAGTCTGCGGGT AAATGA (SEQ ID NO: 75) IgG2 GCCTCCACCACAGCCCCG ASTTAPKVYPLSSCCGDKSSSG X16702 vari- AAAGTCTACCCTCTGAGTT VTLGCLVSSYMPEPVTVTWNS (M25279) ant 3 CTTGCTGCGGGGACAAGT GALKSGVHTFPAVLQSSGLYSL CCAGCTCGGGGGTGACCC SSMVTVPASSSGTQTFTCNVAH TGGGCTGCCTGGTCTCCA PASSTRVDKAVGVSSDCSKPNN GCTACATGCCCGAGCCGG QHCVREPSVFIFPPKPKDTLMI TGACCGTGACCTGGAACT TGTPEVTCVVVNVGHDNPEVQ CGGGTGCCCTGAAGAGCG FSWFVDDVEVHTARTKPREEQ GCGTGCACACCTTCCCGG FNSTYRVVSALPIQHQDWTGG CCGTCCTTCAGTCCTCCG KEFKCKVNIKGLSASIVRIISRS GGCTCTACTCTCTCAGCA KGPAREPQVYVLDPPKEELSKS GCATGGTGACCGTGCCCG TVSLTCMVIGFYPEDVDVEWQ CCAGCAGCTCAGGAACCC RDRQTESEDKYRTTPPQLDAD AGACCTTCACCTGCAACG RSYFLYSKLRVDRNSWQRGDT TAGCCCACCCGGCCAGCA YTCVVMHEALHNHYMQKSTS GCACCAAGGTGGACAAGG KSAGK* CTGTTGGGGTCTCCAGTG (SEQ ID NO: 76) ACTGCTCCAAGCCTAATAA CCAGCATTGCGTGAGGGA ACCATCTGTCTTCATCTTC CCACCGAAACCCAAAGAC ACCCTGATGATCACAGGA ACGCCCGAGGTCACGTGT GTGGTGGTGAACGTGGGC CACGATAACCCCGAGGTG CAGTTCTCCTGGTTCGTG GACGACGTGGAGGTGCAC ACGGCCAGGACGAAGCCG AGAGAGGAGCAGTTCAAC AGCACGTACCGCGTGGTC AGCGCCCTGCCCATCCAG CACCAGGACTGGACTGGA GGAAAGGAGTTCAAGTGC AAGGTCAACATCAAAGGC CTCTCGGCCTCCATCGTG AGGATCATCTCCAGGAGC AAAGGGCCGGCCCGGGAG CCGCAGGTGTATGTCCTG GACCCACCCAAGGAAGAG CTCAGCAAAAGCACGGTC AGCCTCACCTGCATGGTC ATCGGCTTCTACCCAGAA GATGTAGACGTGGAGTGG CAGAGAGACCGGCAGACT GAGTCGGAGGACAAGTAC CGCACGACCCCGCCCCAG CTGGACGCCGACCGCTCC TACTTCCTGTACAGCAAGC TCAGGGTGGACAGGAACA GCTGGCAGAGAGGAGACA CCTACACGTGTGTGGTGA TGCACGAGGCCCTGCACA ATCACTACATGCAGAAGT CCACCTCTAAGTCTGCGG GTAAATGA (SEQ ID NO: 77) IgG3 GCCTCCACCACAGCCCCG ASTTAPKVYPLASSCGDTSSST U63638 vari- AAAGTCTACCCTCTGGCAT VTLGCLVSSYMPEPVTVTWNS ant 1 CCAGCTGCGGAGACACAT GALKSGVHTFPAVRQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPASSSETQTFTCNVAH TGGGCTGCCTGGTCTCCA PASSTKVDKAVTARRPVPTTPK GCTACATGCCCGAGCCGG TTIPPGKPTTPKSEVEKTPCQC TGACCGTGACCTGGAACT SKCPEPLGGLSVFIFPPKPKDT CGGGTGCCCTGAAGAGCG LTISGTPEVFCVVVDVGQDDPE GCGTGCACACCTTCCCGG VQFSWFVDDVEVHTARTKPRE CCGTCCGGCAGTCCTCTG EQFNSTYRVVSALRIQHQDWL GGCTGTACTCTCTCAGCA QGKEFKCKVNNKGLPAPIVRTI GCATGGTGACTGTGCCCG SRTKGQAREPQVYVLAPPREEL CCAGCAGCTCAGAAACCC SKSTLSLTCLITGFYPEEIDVEW AGACCTTCACCTGCAACG QRNGQPESEDKYHTTAPQLDA TAGCCCACCCGGCCAGCA DGSYFLYSKLRVNKSSWQEGD GCACCAAGGTGGACAAGG HYTCAVMHEALRNHYKEKSIS CTGTCACTGCAAGGCGTC RSPGK* CAGTCCCGACGACGCCAA (SEQ ID NO: 78) AGACAACTATCCCTCCTG GAAAACCCACAACCCCAA AGTCTGAAGTTGAAAAGA CACCCTGCCAGTGTTCCA AATGCCCAGAACCTCTGG GAGGACTGTCTGTCTTCAT CTTCCCACCGAAACCCAA GGACACCCTCACAATCTC GGGAACGCCCGAGGTCAC GTGTGTGGTGGTGGACGT GGGCCAGGATGACCCCGA GGTGCAGTTCTCCTGGTT CGTGGACGACGTGGAGGT GCACACGGCCAGGACGAA GCCGAGAGAGGAGCAGTT CAACAGCACCTACCGCGT GGTCAGCGCCCTGCGCAT CCAGCACCAGGACTGGCT GCAGGGAAAGGAGTTCAA GTGCAAGGTCAACAACAA AGGCCTCCCGGCCCCCAT TGTGAGGACCATCTCCAG GACCAAAGGGCAGGCCCG GGAGCCGCAGGTGTATGT CCTGGCCCCACCCCGGGA AGAGCTCAGCAAAAGCAC GCTCAGCCTCACCTGCCT GATCACCGGTTTCTACCCA GAAGAGATAGACGTGGAG TGGCAGAGAAATGGGCAG CCTGAGTCGGAGGACAAG TACCACACGACCGCACCC CAGCTGGATGCTGACGGC TCCTACTTCCTGTACAGCA AGCTCAGGGTGAACAAGA GCAGCTGGCAGGAAGGAG ACCACTACACGTGTGCAG TGATGCACGAAGCTTTAC GGAATCACTACAAAGAGA AGTCCATCTCGAGGTCTC CGGGTAAATGA (SEQ ID NO: 79) IgG3 GCCTCCACCACAGCCCCG ASTTAPKVYPLASRCGDTSSST U63639 vari- AAAGTCTACCCTCTGGCAT VTLGCLVSSYMPEPVTVTWNS ant 2 CCCGCTGCCGAGACACAT GALKSGVHTFPAVLQSSGLYSL CCAGCTCCACCGTGACCC SSMVTVPASTSETQTFTCNVAH TGGGCTGCCTGGTCTCCA PASSTKVDKAVTARRPVPTTPK GCTACATGCCCGAGCCGG TTIPPGKPTTQESEVEKTPCQC TGACCGTGACCTGGAACT SKCPEPLGGLSVFIFPPKPKDT CGGGTGCCCTGAAGAGTG LTISGTPEVTCVVVDVGQDDPE GCGTGCACACCTTCCCGG VQFSWFVDDVEVHTARTKPRE CCGTCCTTCAGTCCTCCG EQFNSTYRVVSALRIQHQDWL GGCTGTACTCTCTCAGCA QGKEFKCKVNNKGLPAPIVRTI GCATGGTGACCGTGCCCG SRTKGQAREPQVYVLAPPREEL CCAGCACCTCAGAAACCC SKSTLSLTCLITGFYPEEIDVEW AGACCTTCACCTGCAACG QRNGQPESEDKYHTTAPQLDA TAGCCCACCCGGCCAGCA DGSYFLYSRLRVNKSSWQEGD GCACCAAGGTGGACAAGG HYTCAVMHEALRNHYKEKSIS CTGTCACTGCAAGGCGTC RSPGK* CAGTCCCGACGACGCCAA (SEQ ID NO: 80) AGACAACCATCCCTCCTG GAAAACCCACAACCCAGG AGTCTGAAGTTGAAAAGA CACCCTGCCAGTGTTCCA AATGCCCAGAACCTCTGG GAGGACTGTCTGTCTTCAT CTTCCCACCGAAACCCAA GGACACCCTCACAATCTC GGGAACGCCCGAGGTCAC GTGTGTGGTGGTGGACGT GGGCCAGGATGACCCCGA GGTGCAGTTCTCCTGGTT CGTGGACGACGTGGAGGT GCACACGGCCAGGACGAA GCCGAGAGAGGAGCAGTT CAACAGCACCTACCGCGT GGTCAGCGCCCTGCGCAT CCAGCACCAGGACTGGCT GCAGGGAAAGGAGTTCAA GTGCAAGGTCAACAACAA AGGCCTCCCGGCCCCCAT TGTGAGGACCATCTCCAG GACCAAAGGGCAGGCCCG GGAGCCGCAGGTGTATGT CCTGGCCCCACCCCGGGA AGAGCTCAGCAAAAGCAC GCTCAGCCTCACCTGCCT GATCACCGGTTTCTACCCA GAAGAGATAGACGTGGAG TGGCAGAGAAATGGGCAG CCTGAGTCGGAGGACAAG TACCACACGACCGCACCC CAGCTGGATGCTGACGGC TCCTACTTCCTGTACAGCA GGCTCAGGGTGAACAAGA GCAGCTGGCAGGAAGGAG ACCACTACACGTGTGCAG TGATGCATGAAGCTTTAC GGAATCACTACAAAGAGA AGTCCATCTCGAGGTCTC CGGGTAAATGA (SEQ ID NO: 81) Bovine Ig CAGCCCAAGTCCCCACCC QPKSPPSVTLFPPSTEELNGNK X62917 Not Chen L. Ig lambda TCGGTCACCCTGTTCCCG ATLVCLISDFYPGSVTVVWKAD regis- et al.,  light CCCTCCACGGAGGAGCTC GSTITRNVETTRASKQSNSKYA tered Vet. chain AACGGCAACAAGGCCACC ASSYLSLTSSDWKSKGSYSCEV Immunol. con- CTGGTGTGTCTCATCAGC THEGSTVTKTVKPSECS* Immuno- stant GACTTCTACCCGGGTAGC (SEQ ID NO: 64) pathol., region GTGACCGTGGTCTGGAAG 124, (CL) GCAGACGGCAGCACCATC 284-294 ACCCGCAACGTGGAGACC (2008). ACCCGGGCCTCCAAACAG PMID: AGCAACAGCAAGTACGCG 18538861 GCCAGCAGCTACCTGAGC CTGACGAGCAGCGACTGG AAATCGAAAGGCAGTTAC AGCTGCGAGGTCACGCAC GAGGGGAGCACCGTGACG AAGACAGTGAAGCCCTCA GAGTGTTCTTAG (SEQ ID NO: 65) Canine Canine IgG-D GCCTCCACCACGGCCCCCTCG ASTTAPSVFPLAPSCGSTS AF35426 http:// Tang L. (Scien- Ig GTTTTCCCACTGGCCCCCAGC GSTVALACLVSGYFPEPVT 7 www. et al., tific heavy TGCGGGTCCACTTCCGGCTCC VSWNSGSLTSGVHTFPSVL imgt.org/ Vet. Name: chain ACGGTGGCCCTGGCCTGCCT QSSGLYSLSSTVTVPSSRW IMGT Immunol. Canis con- GGTGTCAGGCTACTTCCCCGA PSETFTCNVVHPASNTKVD reper Immuno- lupus stant GCCTGTAACTGTGTCCTGGAA KPVPKESTCKCISPCPVPE toire/ pathol. famili- region TTCCGGCTCCTTGACCAGCGG SLGGPSVFIFPPKPKDILRI index.php? 80  aris) (CH1~ TGTGCACACCTTCCCGTCCGT TRTPEITCVVLDLGREDPE section = (3-4), CH3) CCTGCAGTCCTCAGGGCTCTA VQISWFVDGKEVHTAKTQ Locus 259-270 CTCCCTCAGCAGCACGGTGAC PREQQFNSTYRVVSVLPIE Genes &  (2001). AGTGCCCTCCAGCAGGTGGC HQDWLTGKEFKCRVNHIG reper PMID: CCAGCGAGACCTTCACCTGCA LPSPIERTISKARGQAHQPS toire = 1145747 ACGTGGTCCACCCGGCCAGC VYVLPPSPKELSSSDTVTL gene 9 AACACTAAAGTAGACAAGCCA TCLIKDFFPPEIDVEWQSN table & GTGCCCAAAGAGTCCACCTGC GQPEPESKYHTTAPQLDE species = AAGTGTATATCCCCATGCCCA DGSTFLYSKLSVDKSRWQ dog & GTCCCTGAATCACTGGGAGG QGDTFTCAVMHEALQNHY group = GCCTTCGGTCTTCATCTTTCC TDLSLSHSPGK* IGHC CCCGAAACCCAAGGACATCCT (SEQ ID NO: 84) CAGGATTACCCGAACACCCGA GATCACCTGTGTGGTGTTAGA TCTGGGCCGTGAGGACCCTG AGGTGCAGATCAGCTGGTTCG TGGATGGTAAGGAGGTGCAC ACAGCCAAGACGCAGCCTCGT GAGCAGCAGTTCAACAGCACC TACCGTGTGGTCAGCGTCCTC CCCATTGAGCACCAGGACTGG CTCACCGGAAAGGAGTTCAAG TGCAGAGTCAACCACATAGGC CTCCCGTCCCCCATCGAGAGG ACTATCTCCAAAGCCAGAGGG CAAGCCCATCAGCCCAGTGTG TATGTCCTGCCACCATCCCCA AAGGAGTTGTCATCCAGTGAC ACGGTCACCCTGACCTGCCTG ATCAAAGACTTCTTCCCACCT GAGATTGATGTGGAGTGGCA GAGCAATGGACAGCCGGAGC CCGAGAGCAAGTACCACACG ACTGCGCCCCAGCTGGACGA GGACGGGTCCTACTTCCTGTA CAGCAAGCTCTCTGTGGACAA GAGCCGCTGGCAGCAGGGAG ACACCTTCACATGTGCGGTGA TGCATGAAGCTCTACAGAACC ACTACACAGATCTATCCCTCT CCCATTCTCCGGGTAAATGA (SEQ ID NO: 85) Canine Ig CAGCCCAAGGCCTCCCCCTCG QPKASPSVTLFPPSSEELG E02824 Not None Ig lambda GTCACACTCTTCCCGCCCTCC ANKATLVCLISDFYPSGVT regis- light (CL) TCTGAGGAGCTCGGCGCCAA VAWKASGSPVTQGVETTK tered chain CAAGGCCACCCTGGTGTGCCT PSKQSNNKYAASSYLSLTP con- CATCAGCGACTTCTACCCCAG DKWKSHSSFSCLVTHEGS stant CGGCGTGACGGTGGCCTGGA TVEKKVAPAECS* region AGGCAAGCGGCAGCCCCGTC (SEQ ID NO: 82) ACCCAGGGCGTGGAGACCAC CAAGCCCTCCAAGCAGAGCAA CAACAAGTACGCGGCCAGCA GCTACCTGAGCCTGACGCCTG ACAAGTGGAAATCTCACAGCA GCTTCAGCTGCCTGGTCACGC ACGAGGGGAGCACCGTGGAG AAGAAGGTGGCCCCCGCAGA GTGCTCTTAG (SEQ ID NO: 84) Ovine Ovine IgG1 GCCTCAACAACACCCCCGAAA ASTTPPKVYPLTSCCGDTS X69797 http:// Dufour (Scien- Ig GTCTACCCTCTGACTTCTTGC SSIVTLGCLVSSYMPEPVT www. V. et  tific heavy TGCGGGGACACGTCCAGCTC VTWNSGALTSGVHTFPAIL imgt.org/ al., J. Name: chain CATCGTGACCCTGGGCTGCCT QSSGLYSLSSVVTVPASTS IMGT Immun- Ovis con- GGTCTCCAGCTATATGCCCGA GAQTFICNVAHPASSTKVD reper ol., aries) stant GCCGGTGACCGTGACCTGGA KRVEPGCPDPCKHCRCPP toire/ 156, region ACTCTGGTGCCCTGACCAGCG PELPGGPSVFIFPPKPKDT index.php? 2163- (CH1~ GCGTGCACACCTTCCCGGCCA LTISGTPEVTCVVVDVGQD section =  2170 CH3) TCCTGCAGTCCTCCGGGCTCT DPEVQFSWFVDNVEVRTA Locus (1996). ACTCTCTCAGCAGCGTGGTGA RTKPREEQFNSTFRVVSAL Genes & PMID: CCGTGCCGGCCAGCACCTCA PIQHQDWTGGKEFKCKVH reper 8690905 GGAGCCCAGACCTTCATCTGC NEALPAPIVRTISRTKGQA toire = AACGTAGCCCACCCGGCCAG REPQVYVLAPPQEELSKST gene CAGCACCAAGGTGGACAAGC LSVTCLVTGFYPDYIAVEW table & GTGTTGAGCCCGGATGCCCG QKNGQPESEDKYGTTTSQ species = GACCCATGCAAACATTGCCGA LDADGSYFLYSRLRVDKNS sheep & TGCCCACCCCCTGAGCTCCCC WQEGDTYACVVMHEALH group = GGAGGACCGTCTGTCTTCATC NHYTQKSISKPPGK* IGHC TTCCCACCGAAACCCAAGGAC (SEQ ID NO: 90) ACCCTTACAATCTCTGGAACG CCCGAGGTCACGTGTGTGGT GGTGGACGTGGGCCAGGATG ACCCCGAGGTGCAGTTCTCCT GGTTCGTGGACAACGTGGAG GTGCGCACGGCCAGGACAAA GCCGAGAGAGGAGCAGTTCA ACAGCACCTTCCGCGTGGTCA GCGCCCTGCCCATCCAGCACC AAGACTGGACTGGAGGAAAG GAGTTCAAGTGCAAGGTCCAC AACGAAGCCCTCCCGGCCCCC ATCGTGAGGACCATCTCCAGG ACCAAAGGGCAGGCCCGGGA GCCGCAGGTGTACGTCCTGG CCCCACCCCAGGAAGAGCTCA GCAAAAGCACGCTCAGCGTCA CCTGCCTGGTCACCGGCTTCT ACCCAGACTACATCGCCGTGG AGTGGCAGAAAAATGGGCAG CCTGAGTCGGAGGACAAGTA CGGCACGACCACATCCCAGCT GGACGCCGACGGCTCCTACTT CCTGTACAGCAGGCTCAGGGT GGACAAGAACAGCTGGCAAG AAGGAGACACCTACGCGTGT GTGGTGATGCACGAGGCTCT GCACAACCACTACACACAGAA GTCGATCTCTAAGCCTCCGGG TAAATGA (SEQ ID NO: 91) IgG2 GCCTCCACCACAGCCCCGAAA ASTTAPKVYPLTSCCGDTS X70983 Clarkson GTCTACCCTCTGACTTCTTGC SSSSIVTLGCLVSSYMPEPV C. A. et TGCGGGGACACGTCCAGCTC TVTWNSGALTSGVHTFPAI al., Mol. CAGCTCCATCGTGACCCTGGG LQSSGLYSLSSVVTVPASTS Immunol., CTGCCTGGTCTCCAGCTATAT GAQTFICNVAHPASSAKVD 30, GCCCGAGCCGGTGACCGTGA KRVGISSDYSKCSKPPCVS 1195-1204 CCTGGAACTCTGGTGCCCTGA RPSVFIFPPKPKDSLMITGT (1993). CCAGCGGCGTGCACACCTTCC PEVTCVVVDVGQGDPEVQ PMID: CGGCCATCCTGCAGTCCTCCG FSWFVDNVEVRTARTKPR 8413324 GGCTCTACTCTCTCAGCAGCG EEQFNSTFRVVSALPIQHD TGGTGACCGTGCCGGCCAGC HWTGGKEFKCKVHSKGLP ACCTCAGGAGCCCAGACCTTC APIVRTISRAKGQAREPQV ATCTGCAACGTAGCCCACCCG YVLAPPQEELSKSTLSVTC GCCAGCAGCGCCAAGGTGGA LVTGFYPDYIAVEWQRARQ CAAGCGTGTTGGGATCTCCAG PESEDKYGTTTSQLDADGS TGACTACTCCAAGTGTTCTAA YFLYSRLRVDKSSWQRGD ACCGCCTTGCGTGAGCCGACC TYACVVMHEALHNHYTQK GTCTGTCTTCATCTTCCCCCC SISKPPGK* GAAACCCAAGGACAGCCTCAT (SEQ ID NO: 92) GATCACAGGAACGCCCGAGG TCACGTGTGTGGTGGTGGAC GTGGGCCAGGGTGACCCCGA GGTGCAGTTCTCCTGGTTCGT GGACAACGTGGAGGTGCGCA CGGCCAGGACAAAGCCGAGA GAGGAGCAGTTCAACAGCAC CTTCCGCGTGGTCAGCGCCCT GCCCATCCAGCACGACCACTG GACTGGAGGAAAGGAGTTCA AGTGCAAGGTCCACAGCAAA GGCCTCCCGGCCCCCATCGTG AGGACCATCTCCAGGGCCAAA GGGCAGGCCCGGGAGCCGCA GGTGTACGTCCTGGCCCCACC CCAGGAAGAGCTCAGCAAAA GCACGCTCAGCGTCACCTGCC TGGTCACCGGCTTCTACCCAG ACTACATCGCCGTGGAGTGGC AGAGAGCGCGGCAGCCTGAG TCGGAGGACAAGTACGGCAC GACCACATCCCAGCTGGACGC CGACGGCTCCTACTTCCTGTA CAGCAGGCTCAGGGTGGACA AGAGCAGCTGGCAAAGAGGA GACACCTACGCGTGTGTGGTG ATGCACGAGGCTCTGCACAAC CACTACACACAGAAGTCGATC TCTAAGCCTCCGGGTAAATGA (SEQ ID NO: 93) Ovine Ig CCATCCGTCTTCCTCTTCAAA PSVFLFKPSEEQLRTGTVS X54110 Not Jenne Ig kappa CCATCTGAGGAACAGCTGAG VVCLVNDFYPKDINVKVK regis- C. N. et light (CK) GACCGGAACTGTCTCTGTCGT VDGVTQNSNFQNSFTDQD tered al.,  chain GTGCTTGGTGAATGATTTCTA SKKSTYSLSSTLTLSSSEYQ Dev. con- CCCCAAAGATATCAATGTCAA SHNAYACEVSHKSLPTALV Comp. stant GGTGAAAGTGGATGGGGTTA KSFNKNEC* Immunol.  region CCCAGAACAGCAACTTCCAGA (SEQ ID NO: 86) 30 ACAGCTTCACAGACCAGGACA (1-2), GCAAGAAAAGCACCTACAGCC 165-174 TCAGCAGCACCCTGACACTGT (2006). CCAGCTCAGAGTACCAGAGCC PMID: ATAACGCCTATGCGTGTGAGG 1608395 TCAGCCACAAGAGCCTGCCCA 8 CCGCCCTCGTCAAGAGCTTCA ATAAGAATGAATGTTAG (SEQ ID NO: 87) Ig GGTCAGCCCAAGTCCGCACCC GQPKSAPSVTLFPPSTEEL AY73468 lambda TCGGTCACCCTGTTCCCGCCT STNKATVVCLINDFYPGSV 1 (CL) TCCACGGAGGAGCTCAGTACC NVVWKADGSTINQNVKTT AACAAGGCCACCGTGGTGTGT QASKQSNSKYAASSYLTLT CTCATCAACGACTTCTACCCG GSEWKSKSSYTCEVTHEG GGTAGCGTGAACGTGGTCTG STVTKTVKPSECS* GAAGGCAGATGGCAGCACCA (SEQ ID NO: 88) TCAATCAGAACGTGAAGACCA CCCAGGCCTCCAAACAGAGCA ACAGCAAGTACGCGGCCAGC AGCTACCTGACCCTGACGGGC AGCGAGTGGAAGTCTAAGAG CAGTTACACCTGCGAGGTCAC GCACGAGGGGAGCACCGTGA CGAAGACAGTGAAGCCCTCA GAGTGTTCTTAG (SEQ ID NO: 89) Porcine Porcine IgG1^(a) GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCGRDTS U03781 http:// Butler (Scien- Ig GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT www. J. E. et tific heavy GCGGCAGGGACACGTCTGG MTWNSGALTSGVHTFPSV imgt.org/ al., Name: chain CCCTAACGTGGCCTTGGGCT LQPSGLYSLSSMVTVPASS IMGT Immuno- Sus con- GCCTGGCCTCAAGCTACTTC LSSKSYTCNVNHPATTTKV reper genetics scrofa) stant CCCGAGCCAGTGACCATGAC DKRVGTKTKPPCPICPGCE toire/ 61(3): region CTGGAACTCGGGCGCCCTGA VAGPSVFIFPPKPKDTLMIS index.php? 209-230 (CH1~ CCAGTGGCGTGCATACCTTC QTPEVTCVVVDVSKEHAE section = (2009). CH3) CCATCCGTCCTGCAGCCGTC VQFSWYVDGVEVHTAETR Locus PMID: AGGGCTCTACTCCCTCAGCA PKEEQFNSTYRVVSVLPIQ Genes & 1904824 GCATGGTGACCGTGCCGGCC HQDWLKGKEFKCKVNNV reper 8 AGCAGCCTGTCCAGCAAGAG DLPAPITRTISKAIGQSREP toire = Kacsko- CTACACCTGCAATGTCAACC QVYTLPPPAEELSRSKVTV gene vics ACCCGGCCACCACCACCAAG TCLVIGFYPPDIHVEWKSN table & I. et GTGGACAAGCGTGTTGGAAC GQPEPEGNYRTTPPQQDV species = al., J. AAAGACCAAACCACCATGTC DGTFFLYSKLAVDKARWD Pig & Immunol. CCATATGCCCAGGCTGTGAA HGETFECAVMHEALHNHY group = 153(8): GTGGCCGGGCCCTCGGTCTT TQKSISKTQGK* IGHC 3565- CATCTTCCCTCCAAAACCCA (SEQ ID NO: 94) 3573 AGGACACCCTCATGATCTCC (1994). CAGACCCCCGAGGTCACGTG PMID: CGTGGTGGTGGACGTCAGCA 7930579 AGGAGCACGCCGAGGTCCA GTTCTCCTGGTACGTGGACG GCGTAGAGGTGCACACGGC CGAGACGAGACCAAAGGAG GAGCAGTTCAACAGCACCTA CCGTGTGGTCAGCGTCCTGC CCATCCAGCACCAGGACTGG CTGAAGGGGAAGGAGTTCAA GTGCAAGGTCAACAACGTAG ACCTCCCAGCCCCCATCACG AGGACCATCTCCAAGGCTAT AGGGCAGAGCCGGGAGCCG CAGGTGTACACCCTGCCCCC ACCCGCCGAGGAGCTGTCCA GGAGCAAAGTCACCGTAACC TGCCTGGTCATTGGCTTCTA CCCACCTGACATCCATGTTG AGTGGAAGAGCAACGGACA GCCGGAGCCAGAGGGCAAT TACCGCACCACCCCGCCCCA GCAGGACGTGGACGGGACC TTCTTCCTGTACAGCAAGCT CGCGGTGGACAAGGCAAGA TGGGACCATGGAGAAACATT TGAGTGTGCGGTGATGCACG AGGCTCTGCACAACCACTAC ACCCAGAAGTCCATCTCCAA GACTCAGGGTAAATGA (SEQ ID NO: 95) IgG1^(b) GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCGRDVS U03778 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT GCGGCAGGGACGTGTCTGG VTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPASSL GCCTGGCCTCAAGCTACTTC SSKSYTCNVNHPATTTKVD CCCGAGCCAGTGACCGTGAC KRVGIHQPQTCPICPGCEV CTGGAACTCGGGCGCCCTGA AGPSVFIFPPKPKDTLMIS CCAGTGGCGTGCACACCTTC QTPEVTCVVVDVSKEHAE CCATCCGTCCTGCAGCCGTC VQFSWYVDGVEVHTAETR AGGGCTCTACTCCCTCAGCA PKEEQFNSTYRVVSVLPIQ GCATGGTGACCGTGCCGGCC HQDWLKGKEFKCKVNNV AGCAGCCTGTCCAGCAAGAG DLPAPITRTISKAIGQSREP CTACACCTGCAATGTCAACC QVYTLPPPAEELSRSKVTL ACCCGGCCACCACCACCAAG TCLVIGFYPPDIHVEWKSN GTGGACAAGCGTGTTGGAAT GQPEPENTYRTTPPQQDV ACACCAGCCGCAAACATGTC DGTFFLYSKLAVDKARWD CCATATGCCCAGGCTGTGAA HGDKFECAVMHEALHNH GTGGCCGGGCCCTCGGTCTT YTQKSISKTQGK* CATCTTCCCTCCAAAACCCA (SEQ ID NO: 96) AGGACACCCTCATGATCTCC CAGACCCCCGAGGTCACGTG CGTGGTGGTGGACGTCAGCA AGGAGCACGCCGAGGTCCA GTTCTCCTGGTACGTGGACG GCGTAGAGGTGCACACGGC CGAGACGAGACCAAAGGAG GAGCAGTTCAACAGCACCTA CCGTGTGGTCAGCGTCCTGC CCATCCAGCACCAGGACTGG CTGAAGGGGAAGGAGTTCAA GTGCAAGGTCAACAACGTAG ACCTCCCAGCCCCCATCACG AGGACCATCTCCAAGGCTAT AGGGCAGAGCCGGGAGCCG CAGGTGTACACCCTGCCCCC ACCCGCCGAGGAGCTGTCCA GGAGCAAAGTCACGCTAACC TGCCTGGTCATTGGCTTCTA CCCACCTGACATCCATGTTG AGTGGAAGAGCAACGGACA GCCGGAGCCAGAGAACACAT ACCGCACCACCCCGCCCCAG CAGGACGTGGACGGGACCTT CTTCCTGTACAGGAAACTCG CGGTGGACAAGGCAAGATG GGACCATGGAGACAAATTTG AGTGTGCGGTGATGCACGAG GCTCTGCACAACCACTACAC CCAGAAGTCCATCTCCAAGA CTCAGGGTAAATGA (SEQ ID NO: 97) IgG2^(a) GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCSRDTS U03779 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT GCAGGAGGGACACGTCTGG VTWNSGALSSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPASSL GCCTGGCCTCAAGCTACTTC SSKSYTCNVNHPATTTKVD CCCGAGCCAGTGACCGTGAC KRVGTKTKPPCPICPACES CTGGAACTCGGGCGCCCTGT PGPSVFIFPPKPKDTLMISR CCAGTGGCGTGCATACCTTC TPQVTCVVVDVSQENPEV CCATCCGTCCTGCAGCCGTC QFSWYVDGVEVHTAQTRP AGGGCTCTACTCCCTCAGCA KEEQFNSTYRVVSVLPIQH GCATGGTGACCGTGCCGGCC QDWLNGKEFKCKVNNKD AGCAGCCTGTCCAGCAAGAG LPAPITRIISKAKGQTREPQ CTACACCTGCAATGTCAACC VYTLPPHAEELSRSKVSIT ACCCGGCCACCACCACGAAG CLVIGFYPPDIDVEWQRNG GTGGACAAGCGTGTTGGAAC QPEPEGNYRTTPPQQDVD AAAGACCAAACCACCATGTC GTYFLYSKFSVDKASWQG CCATATGCCCAGCCTGTGAA GGIFQCAVMHEALHNHYT TCACCAGGGCCCTCGGTCTT QKSISKTPGK* CATCTTCCCTCCAAAACCCA (SEQ ID NO: 98) AGGACACCCTCATGATCTCC CGGACACCCCAGGTCACGTG CGTGGTGGTTGATGTGAGCC AGGAGAACCCGGAGGTCCA GTTCTCCTGGTACGTGGACG GCGTAGAGGTGCACACGGC CCAGACGAGGCCAAAGGAG GAGCAGTTCAACAGCACCTA CCGCGTGGTCAGCGTCCTAC CCATCCAGCACCAGGACTGG CTGAACGGGAAGGAGTTCAA GTGCAAGGTCAACAACAAAG ACCTCCCAGCCCCCATCACA AGGATCATCTCCAAGGCCAA AGGGCAGACCCGGGAGCCG CAGGTGTACACCCTGCCCCC ACACGCCGAGGAGCTGTCCA GGAGCAAAGTCAGCATAACC TGCCTGGTCATTGGCTTCTA CCCACCTGACATCGATGTCG AGTGGCAAAGAAACGGACA GCCGGAGCCAGAGGGCAAT TACCGCACCACCCCGCCCCA GCAGGACGTGGACGGGACC TACTTCCTGTACAGCAAGTT CTCGGTGGACAAGGCCAGCT GGCAGGGTGGAGGCATATTC CAGTGTGCGGTGATGCACGA GGCTCTGCACAACCACTACA CCCAGAAGTCTATCTCCAAG ACTCCGGGTAAATGA (SEQ ID NO: 99) IgG2^(b) GCCCCCAAGACGGCCCCATT APKTAPLVYPLAPCGRDTS U03780 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT GCGGCAGGGACACGTCTGG VTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPASSL GCCTGGCCTCAAGCTACTTC SSKSYTCNVNHPATTTKVD CCCGAGCCAGTGACCGTGAC KRVGTKTKPPCPICPACES CTGGAACTCGGGCGCCCTGA PGPSVFIFPPKPKDTLMISR CCAGTGGCGTGCATACCTTC TPQVTCVVVDVSQENPEV CCATCCGTCCTGCAGCCGTC QFSWYVDGVEVHTAQTRP AGGGCTCTACTCCCTCAGCA KEEQFNSTYRVVSVLPIQH GCATGGTGACCGTGCCGGCC QDWLNGKEFKCKVNNKD AGCAGCCTGTCCAGCAAGAG LPAPITRIISKAKGQTREPQ CTACACCTGCAATGTCAACC VYTLPPHAEELSRSKVSIT ACCCGGCCACCACCACCAAG CLVIGFYPPDIDVEWQRNG GTGGACAAGCGTGTTGGAAC QPEPEGNYRTTPPQQDVD AAAGACCAAACCACCATGTC GTYFLYSKFSVDKASWQG CCATATGCCCAGCCTGTGAA GGIFQCAVMHEALHNHYT TCGCCAGGGCCCTCGGTCTT QKSISKTPGK* CATCTTCCCTCCAAAACCCA (SEQ ID NO: 100) AGGACACCCTCATGATCTCC CGGACACCCCAGGTCACGTG CGTGGTAGTTGATGTGAGCC AGGAGAACCCGGAGGTCCA GTTCTCCTGGTACGTGGACG GCGTAGAGGTGCACACGGC CCAGACGAGGCCAAAGGAG GAGCAGTTCAACAGCACCTA CCGCGTGGTCAGCGTCCTGC CCATCCAGCACCAGGACTGG CTGAACGGGAAGGAGTTCAA GTGCAAGGTCAACAACAAAG ACCTCCCAGCCCCCATCACA AGGATCATCTCCAAGGCCAA AGGGCAGACCCGGGAGCCG CAGGTGTACACCCTGCCCCC ACACGCCGAGGAGCTGTCCA GGAGCAAAGTCAGCATAACC TGCCTGGTCATTGGCTTCTA CCCACCTGACATCGATGTCG AGTGGCAAAGAAACGGACA GCCGGAGCCAGAGGGCAAT TACCGCACCACCCCGCCCCA GCAGGACGTGGACGGGACC TACTTCCTGTACAGCAAGTT CTCGGTGGACAAGGCCAGCT GGCAGGGTGGAGGCATATTC CAGTGTGCGGTGATGCACGA GGCTCTGCACAACCACTACA CCCAGAAGTCTATCTCCAAG ACTCCGGGTAAATGA (SEQ ID NO: 01) IgG3 GCCTACAACACAGCTCCATC AYNTAPSVYPLAPCGRDVS EU372658 GGTCTACCCTCTGGCCCCCT DHNVALGCLVSSYFPEPVT GTGGCAGGGACGTGTCTGAT VTWNSGALSRVVHTFPSVL CATAACGTGGCCTTGGGCTG QPSGLYSLSSMVIVAASSLS CCTTGTCTCAAGCTACTTCC TLSYTCNVYHPATNTKVD CCGAGCCAGTGACCGTGACC KRVDIEPPTPICPEICSCPA TGGAACTCGGGTGCCCTGTC AEVLGAPSVFLFPPKPRDI CAGAGTCGTGCATACCTTCC LMISRTPKVTCVVVDVSQE CATCCGTCCTGCAGCCGTCA EAEVQFSWYVDGVQLYTA GGGCTCTACTCCCTCAGCAG QTRPMEEQFNSTYRVVSV CATGGTGATCGTGGCGGCCA LPIQHQDWLKGKEFKCKV GCAGCCTGTCCACCCTGAGC NNKDLLSPITRTISKATGPS TACACGTGCAACGTCTACCA RVPQVYTLPPAWEELSKSK CCCGGCCACCAACACCAAGG VSITCLVTGFYPPDIDVEW TGGACAAGCGTGTTGACATC QSNGQQEPEGNYRTTPPQ GAACCCCCCACACCCATCTG QDVDGTYFLYSKLAVDKV TCCCGAAATTTGCTCATGCC RWQRGDLFQCAVMHEAL CAGCTGCAGAGGTCCTGGGA HNHYTQKSISKTQGK GCACCGTCGGTCTTCCTCTT (SEQ ID NO: 102) CCCTCCAAAACCCAAGGACA TCCTCATGATCTCCCGGACA CCCAAGGTCACGTGCGTGGT GGTGGACGTGAGCCAGGAG GAGGCTGAAGTCCAGTTCTC CTGGTACGTGGACGGCGTAC AGTTGTACACGGCCCAGACG AGGCCAATGGAGGAGCAGTT CAACAGCACCTACCGCGTGG TCAGCGTCCTGCCCATCCAG CACCAGGACTGGCTGAAGG GGAAGGAGTTCAAGTGCAAG GTCAACAACAAAGACCTCCT TTCCCCCATCACGAGGACCA TCTCCAAGGCTACAGGGCCG AGCCGGGTGCCGCAGGTGT ACACCCTGCCCCCAGCCTGG GAAGAGCTGTCCAAGAGCAA AGTCAGCATAACCTGCCTGG TCACTGGCTTCTACCCACCT GACATCGATGTCGAGTGGCA GAGCAACGGACAACAAGAG CCAGAGGGCAATTACCGCAC CACCCCGCCCCAGCAGGACG TGGATGGGACCTACTTCCTG TACAGCAAGCTCGCGGTGGA CAAGGTCAGGTGGCAGCGT GGAGACCTATTCCAGTGTGC GGTGATGCACGAGGCTCTGC ACAACCACTACACCCAGAAG TCCATCTCCAAGACTCAGGG TAAATGA (SEQ ID NO: 103) IgG4a ACCTTCCCATCCGTCCTGCA TFPSVLQPSGLYSLSSMVT U03782 GCCGTCAGGGCTCTACTCCC VPASSLSSKSYTCNVNHPA TCAGCAGGATGGTGACCGTG TTTKVDKRVGTKTKPPCPI CCGGCCAGCAGCCTGTCCAG CPACEGPGPSAFIFPPKPK CAAGAGCTACACCTGCAATG DTLMISRTPKVTCVVVDVS TCAACCACCCGGCCACCACC QENPEVQFSWYVDGVEVH ACCAAGGTGGACAAGCGTGT TAQTRPKEEQFNSTYRVVS TGGAACAAAGACCAAACCAC VLPIQHQDWLNGKEFKCK CATGTCCCATATGCCCAGCC VNNKDLPAPITRIISKAKG TGTGAAGGGCCCGGGCCCTC QTREPQVYTLPPPTEELSR GGCCTTCATCTTCCCTCCAA SKVTLTCLVTGFYPPDIDV AACCCAAGGACACCCTCATG EWQRNGQPEPEGNYRTTP ATCTCCCGGACCCCCAAGGT PQQDVDGTYFLYSKLAVD CACGTGCGTGGTGGTAGATG KASWQRGDTFQCAVMHE TGAGCCAGGAGAACCCGGA ALHNHYTQKSIFKTGK* GGTCCAGTTCTCCTGGTACG (SEQ ID NO: 104) TGGACGGCGTAGAGGTGCA CACGGCCCAGACGAGGCCA AAGGAGGAGCAGTTCAACAG CACCTACCGCGTGGTCAGCG TCCTGCCCATCCAGCACCAG GACTGGCTGAACGGGAAGG AGTTCAAGTGCAAGGTCAAC AACAAAGACCTCCCAGCCCC CATCACAAGGATCATCTCCA AGGCCAAAGGGCAGACCCG GGAGCCGCAGGTGTACACCC TGCCCCCACCCACCGAGGAG CTGTCCAGGAGCAAAGTCAC GCTAACCTGCCTGGTCACTG GCTTCTACCCACCTGACATC GATGTCGAGTGGCAAAGAAA CGGACAGCCGGAGCCAGAG GGCAATTACCGCACCACCCC GCCCCAGCAGGACGTGGAC GGGACCTACTTCCTGTACAG CAAGCTCGCGGTGGACAAG GCCAGCTGGCAGCGTGGAG ACACATTCCAGTGTGCGGTG ATGCACGAGGCTCTGCACAA CCACTACACCCAGAAGTCCA TCTTGAAGACTCCGGGTAAA TGA (SEQ ID NO: 105) IgG4b GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCGRDVS EU372654 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT GCGGCAGGGACGTGTCTGG VTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPASSL GCCTGGCCTCAAGCTACTTC SSKSYTCNVNHPATTTKVD CCCGAGCCAGTGACCGTGAC KRVGIHQPQTCPICPACEG CTGGAACTCGGGCGCCCTGA PGPSAFIFPPKPKDTLMISR CCAGTGGCGTGCACACCTTC TPKVTCVVVDVSQENPEV CCATCCGTCCTGCAGCCGTC QFSWYVDGVEVHTAQTRP AGGGCTCTACTCCCTGAGCA KEEQFNSTYRVVSVLLIQH GCATGGTGACCGTGCCGGCC QDWLNGKEFECKVNNKD AGCAGCCTGTCCAGCAAGAG LPAPITRIISKAKGQTREPQ CTACACCTGCAATGTCAACC VYTLPPPTEELSRSLVTLT ACCCGGCCACCACCACCAAG CLVTGFYPPDIDVEWQRN GTGGACAAGCGTGTTGGAAT GQPEPEGNYRTTPPQQDV ACACCAGCCGCAAACATGTC DGTYFLYSKLAVDKASWQ CCATATGCCCAGCCTGTGAA RGDTFQCAVMHEALHNHY GGGCCCGGGCCCTCGGCCTT T (SEQ ID NO: 106) CATCTTCCCTCCAAAACCCA AGGACACCCTCATGATCTCC CGGACCCCCAAGGTCACGTG CGTGGTGGTTGATGTGAGCC AGGAGAACCCGGAGGTCCA GTTCTCCTGGTACGTGGACG GCGTAGAGGTGCACACGGC CCAGACGAGGCCAAAGGAG GAGCAGTTCAACAGCACCTA CCGCGTGGTCAGCGTCCTGC TCATCCAGCACCAGGACTGG CTGAACGGGAAGGAGTTCAA GTGCAAGGTCAACAACAAAG ACCTCCCAGCCCCCATCACA AGGATCATCTCCAAGGCCAA AGGGCAGACCCGGGAGCCG CAGGTGTACACCCTGCCCCC ACCCACCGAGGAGCTGTCCA GGAGCAAAGTCACGCTAACC TGCCTGGTCACTGGCTTCTA CCCACCTGACATCGATGTCG AGTGGCAAAGAAACGGACA GCCGGAGCCAGAGGGCAAT TACCGCACCACCCCGCCCCA GCAGGACGTGGACGGGACC TACTTCCTGTACAGCAAGCT CGCGGTGGACAAGGCCAGC TGGCAGCGTGGAGACACATT CCAGTGTGCGGTGATGCACG AGGCTCTGCACAACCACTAC ACCC (SEQ ID NO: 107) IgG5a GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCSRDTS EU37265 GGTCTACCCTCTGGCCCCCT GPNVALGCLVSSYFPEPVT 7 GCAGCAGGGACACGTCTGG VTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPAHSL GCCTGGTCTCAAGCTACTTC SSKRYTCNVNHPATKTKV CCCGAGCCAGTGACCGTGAC DLCVGRPCPICPGCEVAGP CTGGAACTCGGGCGCCCTGA SVFIFPPKPKDILMISRTPE CCAGTGGCGTGCACACCTTC VTCVVVDVSKEHAEVQFS CCATCCGTCCTGCAGCCGTC WYVDGEEVHTAETRPKEE AGGGCTCTACTCCCTCAGCA QFNSTYRVVSVLPIQHEDW GCATGGTGACCGTGCCGGCC LKGKEFECKVNNEDLPGP CACAGCTTGTCCAGCAAGCG ITRTISKAKGVVRSPEVYTL CTATACGTGCAATGTCAACC PPPAEELSKSIVTLTCLVKS ACCCAGCCACCAAAACCAAG IFP?FIHVEWKINGKPEPE GTGGACCTGTGTGTTGGACG NAYRTTPPQEDEDRTYFLY ACCATGTCCCATATGCCCAG SKLAVDKARWDHGETFEC GCTGTGAAGTGGCCGGGCC AVMHEALHNHYTQKSISK CTCGGTCTTCATCTTCCCTC TQGK* CAAAACCCAAGGACATCCTC (SEQ ID NO: 108) ATGATCTCCCGGACCCCCGA GGTCACGTGCGTGGTGGTG GACGTCAGCAAGGAGCACG CCGAGGTCCAGTTCTCCTGG TACGTGGACGGCGAAGAGG TGCACACGGCCGAGACGAG GCCAAAGGAGGAGCAGTTCA ACAGCACCTACCGCGTGGTC AGCGTCCTGCCCATCCAGCA CGAGGACTGGCTGAAGGGG AAGGAGTTCGAGTGCAAGGT CAACAACGAAGACCTCCCAG GCCCCATCACGAGGACCATC TCCAAGGCCAAAGGGGTGGT ACGGAGCCCGGAGGTGTAC ACCCTGCCCCCACCCGCCGA GGAGCTGTCCAAGAGCATAG TCACGCTAACCTGCCTGGTC AAAAGCATCTTCCCGNCTTT CATCCATGTTGAGTGGAAAA TCAACGGAAAACCAGAGCCA GAGAACGCATATCGCACCAC CCCGCCTCAGGAGGACGAG GACAGGACCTACTTCCTGTA CAGCAAGCTCGCGGTGGACA AGGCAAGATGGGACCATGG AGAAACATTTGAGTGTGCGG TGATGCACGAGGCTCTGCAC AACCACTACACCCAGAAGTC CATCTCCAAGACTCAGGGTA AATGA (SEQ ID NO: 109) IgG5b GCCTACAACACAGCTCCATC AYNTAPSVYPLAPCGRDVS EU37265 GGTCTACCCTCTGGCCCCCT DHNVALGCLVSSYFPEPVT 6 GTGGCAGGGACGTGTCTGAT VTWNWGAQTSGVHTFPSV CATAACGTGGCCTTGGGCTG LQPSGLYSLSSTVTVPAHS CCTGGTCTCAAGCTACTTCC ISSKCFTCNVNHPATTTKV CCGAGCCAGTGACCGTGACC DLCVGKKTKPRCPICPGCE TGGAACTGGGGCGCCCAGA VAGPSVFIFPPKPKDILMIS CCAGTGGCGTGCACACCTTC RTPEVTCVVVDVSKEHAE CCATCCGTCCTGCAGCCGTC VQFSWYVDGEEVHTAETR AGGGCTCTACTCCCTCAGCA PKEEQFNSTYRVVSVLPIQ GCACGGTGACCGTGCCGGC HEDWLKGKEFECKVNNE CCACAGCTTGTCCAGCAAGT DLPGPITRTISKAKGVVRSP GCTTCACGTGCAATGTCAAC EVYTLPPPAEELSKSIVTLT CACCCGGCCACCACCACCAA CLVKSFFPPFIHVEWKING GGTGGACCTGTGTGTTGGAA KPEPENAYRTTPPQEDED AAAAGACCAAGCCTCGATGT GTYFLYSKFSVEKFRWHS CCCATATGCCCAGGCTGTGA GGIHCAVMHEALHNHYT AGTGGCCGGGCCCTCGGTCT (SEQ ID NO: 110) TCATCTTCCCTCCAAAACCC AAGGACATCCTCATGATCTC CCGGACCCCCGAGGTCACGT GCGTGGTGGTGGACGTCAG CAAGGAGCACGCCGAGGTC CAGTTCTCCTGGTACGTGGA CGGCGAAGAGGTGCACACG GCCGAGACGAGACCAAAGG AGGAGCAGTTCAACAGCACT TACCGCGTGGTCAGCGTCCT GCCCATCCAGCACGAGGACT GGCTGAAGGGGAAGGAGTT CGAGTGCAAGGTCAACAACG AAGACCTCCCAGGCCCCATC ACGAGGACCATCTCCAAGGC CAAAGGGGTGGTACGGAGC CCGGAGGTGTACACCCTGCC CCCACCCGCCGAGGAGCTGT CCAAGAGCATAGTCACGCTA ACCTGCCTGGTCAAAAGCTT CTTCCCGCCTTTCATCCATG TTGAGTGGAAAATCAACGGA AAACCAGAGCCAGAGAACGC ATACCGCACCACCCCGCCCC AGGAGGACGAGGACGGGAC CTACTTCCTGTACAGCAAGT TCTCGGTGGAAAAGTTCAGG TGGCACAGTGGAGGCATCCA CTGTGCGGTGATGCACGAGG CTCTGCACAACCACTACACC C (SEQ ID NO: 111) IgG6a GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCGRDTS EU37265 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT 5 GCGGCAGGGACACGTCTGG LTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSMVTVPASSL GCCTGGCCTCAAGCTACTTC SSKSYTCNVNHPATTTKVD CCCGAGCCAGTGACCCTGAC LCVGRPCPICPACEGPGPS CTGGAACTCGGGCGCCCTGA VFIFPPKPKDTLMISRTPQ CCAGTGGCGTGCATACCTTC VTCVVVDVSQENPEVQFS CCATCCGTCCTGCAGCCGTC WYVDGVEVHTAQTRPKEA AGGGCTCTACTCCCTCAGCA QFNSTYRVVSVLPIQHEDW GCATGGTGACCGTGCCGGCC LKGKEFECKVNNKDLPAP AGCAGCCTGTCCAGCAAGAG ITRIISKAKGPSREPQVYTL CTACACCTGCAATGTCAACC SPSAEELSRSKVSITCLVTG ACCCGGCCACCACCACCAAG FYPPDIDVEWKSNGQPEP GTGGACCTGTGTGTTGGACG EGNYRTTPPQQDVDGTYF ACCATGTCCCATATGCCCAG LYSKLAVDKASWQRGDPF CCTGTGAAGGGCCCGGGCC QCAVMHEALHNHYT CTCGGTCTTCATCTTCCCTC (SEQ ID NO: 112) CAAAACCCAAGGACACCCTC ATGATCTCCCGGACACCCCA GGTCACGTGCGTGGTGGTAG ATGTGAGCCAGGAAAACCCG GAGGTCCAGTTCTCCTGGTA TGTGGACGGTGTAGAGGTGC ACACGGCCCAGACGAGGCC AAAGGAGGCGCAGTTCAACA GCACCTACCGTGTGGTCAGC GTCCTGCCCATCCAGCACGA GGACTGGCTGAAGGGGAAG GAGTTCGAGTGCAAGGTCAA CAACAAAGACCTCCCAGCCC CCATCACAAGGATCATCTCC AAGGCCAAAGGGCCGAGCC GGGAGCCGCAGGTGTACAC CCTGTCCCCATCCGCCGAGG AGCTGTCCAGGAGCAAAGTC AGCATAACCTGCCTGGTCAC TGGCTTCTACCCACCTGACA TCGATGTCGAGTGGAAGAGC AACGGACAGCCGGAGCCAG AGGGCAATTACCGCACCACC CCGCCCCAGCAGGACGTGG ACGGGACCTACTTCCTGTAC AGCAAGCTCGCGGTGGACAA GGCCAGCTGGCAGCGTGGA GACCCATTCCAGTGTGCGGT GATGCACGAGGCTCTGCACA ACCACTACACCC (SEQ ID NO: 113) IgG6b GCCCCCAAGACGGCCCCATC APKTAPSVYPLAPCGRDTS EU37265 GGTCTACCCTCTGGCCCCCT GPNVALGCLASSYFPEPVT 3 GCGGCAGGGACACGTCTGG VTWNSGALTSGVHTFPSVL CCCTAACGTGGCCTTGGGCT QPSGLYSLSSTVTVPARSSS GCCTGGCCTCAAGCTACTTC RKCFTCNVNHPATTTKVD CCCGAGCCAGTGACCGTGAC LCVGRPCPICPACEGNGPS CTGGAACTCGGGCGCCCTGA VFIFPPKPKDTLMISRTPEV CCAGTGGCGTGCACACCTTC TCVVVDVSQENPEVQFSW CCATCCGTCCTGCAGCCGTC YVDGEEVHTAETRPKEEQ AGGGCTCTACTCCCTCAGCA FNSTYRVVSVLPIQHQDWL GCACGGTGACCGTGCCGGC KGKEFECKVNNKDLPAPI CAGGAGCTCGTCCAGAAAGT TRIISKAKGPSREPQVYTLS GCTTCACGTGCAATGTCAAC PSAEELSRSKVSITCLVTGF CACCCGGCCACCACCACCAA YPPDIDVEWKSNGQPEPE GGTGGACCTGTGTGTTGGAC GNYRSTPPQEDEDGTYFLY GACCATGTCCCATATGCCCA SKLAVDKARLQSGGIHCAV GCCTGTGAAGGGAACGGGC MHEALHNHYTQKSISKT CCTCGGTCTTCATCTTCCCT (SEQ ID NO: 114) CCAAAACCCAAGGACACCCT CATGATCTCCCGGACCCCCG AGGTCACGTGCGTGGTGGTA GATGTGAGCCAGGAAAACCC GGAGGTCCAGTTCTCCTGGT ACGTGGACGGCGAAGAGGT GCACACGGCCGAGACGAGG CCAAAGGAGGAGCAGTTCAA CAGCACCTACCGTGTGGTCA GCGTCCTGCCCATCCAGCAC CAGGACTGGCTGAAGGGAA AGGAGTTCGAGTGCAAGGTC AACAACAAAGACCTCCCAGC CCCCATCACAAGGATCATCT CCAAGGCCAAAGGGCCGAG CCGGGAGCCGCAGGTGTAC ACCCTGTCCCCATCCGCCGA GGAGCTGTCCAGGAGCAAA GTCAGCATAACCTGCCTGGT CACTGGCTTCTACCCACCTG ACATCGATGTCGAGTGGAAG AGCAACGGACAGCCGGAGC CAGAGGGCAATTACCGCTCC ACCCCGCCCCAGGAGGACG AGGACGGGACCTACTTCCTG TACAGCAAACTCGCGGTGGA CAAGGCGAGGTTGCAGAGT GGAGGCATCCACTGTGCGGT GATGCACGAGGCTCTGCACA ACCACTACACCCAGAAGTCC ATCTCCAAGACT (SEQ ID NO: 115) Porcine Ig FP31289 http:// Schwartz Ig  kappa 8 www. J. C. et light (CK)  imgt.org/ al.,  chain vari- IMGT Immuno- con- ant 1 reper gene- stant toire/ tics, region index.php? 64, section = 303-311 Locus (2012). Genes & PMID: reper 2210954 toire = 0 gene table & species = Pig & group = IGLC Ig CU69484 11 kappa 8 (CK) vari- ant 2 Ig CU46766 http:// lambda 9 www. (CL) imgt.org/ vari- IMGT ant 1 reper toire/ index.php? section = Locus Genes & reper toire = gene table & species = Pig & group = IGKC Ig CU46759 lambda 9 (CK) vari- ant 2 Water Water IgG1? GAGCGGCGTGCACACCTTCCC SGVHTFPAVLQSSGLYSLS NW_005 Not None buffalo buffalo GGCCGTCCTTCAGTCCTCCGG STVTAPASATKSQTFTCNV 690903 regis- (Scien- Ig GCTCTACTCTCTCAGCAGCAC AHPASSTKVDKAVVPPCRP tered tific heavy GGTGACCGCGCCCGCCAGCG KPCDCCPPPELPGGPSVFI Name: chain CCACAAAAAGCCAGACCTTCA FPPKPKDTLTISGTPEVTC Bubalus con- CCTGCAACGTAGCCCACCCGG VVVDVGHDDPEVKFSWFV bubalis stant CCAGCAGCACCAAGGTGGAC DDVEVNTARTKPREEQFN region AAGGCTGTTGTTCCCCCATGC STYRVVSALPIQHNDWTG (CH1~ AGACCGAAACCCTGTGATTGC GKEFKCKVYNEGLPAPIVR CH3) TGCCCACCCCCTGAGCTCCCC TISRTKGQAREPQVYVLAP GGAGGACCCTCTGTCTTCATC PQDELSKSTVSITCMVTGF TTCCCACCAAAACCCAAGGAC YPDYIAVEWQKDGQPESE ACCCTCACAATCTCTGGAACT DKYGTTPPQLDSDGSYFLY CCTGAGGTCACGTGTGTGGTG SRLRVNKNSWQEGGAYTC GTGGACGTGGGCCACGATGA VVMHE CCCCGAGGTGAAGTTCTCCTG (SEQ ID NO: 118) GTTCGTGGACGATGTGGAGG TAAACACAGCCAGGACGAAG CCAAGAGAGGAGCAGTTCAA CAGCACCTACCGCGTGGTCAG CGCCCTGCCCATCCAGCACAA CGACTGGACTGGAGGAAAGG AGTTCAAGTGCAAGGTCTACA ATGAAGGCCTCCCAGCCCCCA TCGTGAGGACCATCTCCAGGA CCAAAGGGCAGGCCCGGGAG CCGCAGGTGTACGTCCTGGCC CCACCCCAGGACGAGCTCAG CAAAAGCACGGTCAGCATCAC TTGCATGGTCACTGGCTTCTA CCCAGACTACATCGCCGTAGA GTGGCAGAAAGATGGGCAGC CTGAGTCAGAGGACAAATATG GCACGACCCCGCCCCAGCTG GACAGCGATGGCTCCTACTTC CTGTACAGCAGGCTCAGGGT GAACAAGAACAGCTGGCAAG AAGGAGGCGCCTACACGTGT GTAGTGATGCATGAGGC (SEQ ID NO: 119) IgG2? GCCTCCATCACAGCCCCGAAA ASITAPKVYPLTSCRGETSS NW_005 GTCTACCCTCTGACTTCTTGC STVTLGCLVSSYMPEPVTV 766143 CGCGGGGAAACGTCCAGCTC TWNSGALKSGVHTFPAVL CACCGTGACCCTGGGCTGCCT QSSGLYSLSSTVTAPASAT GGTCTCCAGCTACATGCCCGA KSQTFTCNVAHPASSTKVD GCCGGTGACCGTGACCTGGA TAVGFSSDCCKFPKPCVRG ACTCGGGTGCCCTGAAGAGC PSVFIFPPKPKDTLMITGN GGCGTGCACACCTTCCCGGCC PEVTCVVVDVGRDNPEVQ GTCCTTCAGTCCTCTGGGCTC FSWFVGDVEVHTGRSKPR TACTCTCTCAGCAGCACGGTG EEQFNSTYRVVSTLPIQHN ACCGCGCCCGCCAGCGCCAC DWTGGKEFKCKVNNKGL AAAAAGCCAGACCTTCACCTG PAPIVRTISRTKGQAREPQ CAACGTAGCCCACCCGGCCA VYVLAPPQEELSKSTVSVT GCAGCACCAAGGTGGACACG CMVTGFYPDYIAVEWHRD GCTGTTGGGTTCTCCAGTGAC RQAESEDKYRTTPPQLDSD TGCTGCAAGTTTCCTAAGCCT GSYFLYSRLKVNKNSWQE TGTGTGAGGGGACCATCTGTC GGAYTCVVMHE TTCATCTTCCCGCCGAAACCC (SEQ ID NO: 120) AAAGACACCCTGATGATCACA GGAAATCCCGAGGTCACATGT GTGGTGGTGGACGTGGGCCG GGATAACCCCGAGGTGCAGTT CTCCTGGTTCGTGGGTGATGT GGAGGTGCACACGGGCAGGT CGAAGCCGAGAGAGGAGCAG TTCAACAGCACCTACCGCGTG GTCAGCACCCTGCCCATCCAG CACAATGACTGGACTGGAGG AAAGGAGTTCAAGTGCAAGGT CAACAACAAAGGCCTCCCAGC CCCCATCGTGAGGACCATCTC CAGGACCAAAGGGCAGGCCC GGGAGCCGCAGGTGTACGTC CTGGCCCCACCCCAGGAAGA GCTCAGCAAAAGCACGGTCA GCGTCACTTGCATGGTCACTG GCTTCTACCCAGACTACATCG CCGTAGAGTGGCATAGAGAC CGGCAGGCTGAGTCGGAGGA CAAGTACCGCACGACCCCGCC CCAGCTGGACAGCGATGGCT CCTACTTCCTGTACAGCAGGC TCAAGGTGAACAAGAACAGCT GGCAAGAAGGAGGCGCCTAC ACGTGTGTAGTGATGCATGAG GC (SEQ ID NO: 121) IgG3? GCCTCCACCACAGCCCCGAAA ASTTAPKVYPLASSCGDTS NW_005 GTCTACCCTCTGGCATCCAGC SSTVTLGCLVSSYMPEPVT 784206 TGCGGGGACACGTCCAGCTC VTWNSGALKNGVHTFPAV CACCGTGACCCTGGGCTGCCT RQSSGLYSLSSMVTMPTST GGTCTCCAGCTACATGCCCGA AGTQTFTCNVAHPASSTKV GCCGGTGACCGTGACCTGGA DTAVTARHPVPKTPETPIH ACTCGGGTGCCCTGAAGAAC PVKPPTQEPRDEKTPCQCP GGCGTGCACACCTTCCCGGCC KCPEPLGGLSVFIFPPKPK GTCCGGCAGTCCTCCGGGCTC DTLTISGTPEVTCVVVDVG TACTCTCTCAGCAGCATGGTG QDDPEVQFSWFVDDVEVH ACCATGCCCACCAGCACCGCA TARMKPREEQFNSTYRVV GGAACCCAGACCTTCACCTGC SALPIQHQDWLREKEFKC AACGTAGCCCACCCGGCCAG KVNNKGLPAPIVRTISRTK CAGCACCAAGGTGGACACGG GQAREPQVYVLAPPREELS CTGTCACTGCAAGGCATCCGG KSTLSLTCLITGFYPEEVD TCCCGAAGACACCAGAGACAC VEWQRNGQPESEDKYHTT CTATCCATCCTGTAAAACCCC PPQLDADGSYFLYSRLRVN CAACCCAGGAGCCCAGAGAT RSSWQEGDHYTCAVMHEA GAAAAGACACCCTGCCAGTGT LRNHYKEKPISRSPGK* CCCAAATGCCCAGAACCTCTG (SEQ ID NO: 122) GGAGGACTGTCTGTCTTCATC TTCCCACCGAAACCCAAGGAC ACCCTCACAATCTCTGGAACG CCCGAGGTCACGTGTGTGGT GGTCGACGTGGGCCAGGATG ACCCCGAAGTGCAGTTCTCCT GGTTCGTGGATGACGTGGAG GTGCACAGAGCCAGGATGAA GCCAAGAGAGGAGCAGTTCA ACAGCACCTACCGCGTGGTCA GCGCCCTGCCCATCCAGCACC AGGACTGGCTGCGGGAAAAG GAGTTCAAGTGCAAGGTCAAC AACAAAGGCCTCCCGGCCCCC ATCGTGAGGACCATCTCCAGG ACCAAAGGGCAGGCCCGGGA GCCACAGGTGTATGTCCTGGC CCCACCCCGGGAAGAGCTCA GCAAAAGCACGCTCAGCCTCA CCTGCCTAATCACCGGCTTCT ACCCAGAAGAGGTAGACGTG GAGTGGCAGAGAAATGGGCA GCCTGAGTCAGAGGACAAGT ACCACACGACCCCACCCCAGC TGGACGCTGACGGCTCCTACT TCCTGTACAGCAGGCTCAGGG TGAACAGGAGCAGCTGGCAG GAAGGAGACCACTACACGTGT GCAGTGATGCATGAAGCTTTA CGGAATCACTACAAAGAGAAG CCCATCTCGAGGTCTCCGGGT AAATGA (SEQ ID NO: 123) Water Ig CAGCCCAAGTCCGCACCCTCA QPKSAPSVTLFPPSTEELS NW_005 Not None buf- lamb- GTCACCCTGTTCCCACCCTCC ANKATLVCLISDFYPGSMT 690786 regis- falo  da? ACGGAGGAGCTCAGCGCCAA VARKADGSTITRNVETTRA tered Ig CAAGGCCACCCTGGTGTGTCT SKQSNSKYAASSYLSLTGS light CATCAGCGACTTCTACCCGGG EWKSKGSYSCEVTHEGST chain TAGCATGACCGTGGCCAGGA VTKTVKPSECS* con- AGGCAGACGGCAGCACCATC (SEQ ID NO: 116) stant ACCCGGAACGTGGAGACCAC region CCGGGCCTCCAAACAGAGCA (CL) ACAGCAAGTACGCGGCCAGC AGCTACCTGAGCCTGACGGG CAGCGAGTGGAAATCGAAAG GCAGTTACAGCTGCGAGGTCA CGCACGAGGGGAGCACCGTG ACAAAGACAGTGAAGCCCTCA GAGTGTTCTTAG (SEQ ID NO: 117)

The amino acid sequences as shown in SEQ ID NOS: 8 to 13, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120 and 122 may have deletion(s), substitution(s) or addition(s) of one or several (e.g., up to five, about 10 at the most) amino acids. Even when such mutations have been introduced, the resulting amino acid sequences are capable of having the function as the constant region of Ig heavy chain or light chain.

The anti-PD-L1 antibody of the present invention may have a four-chain structure comprising two light chains and two heavy chains.

The anti-PD-L1 antibody of the present invention may be prepared as described below. Briefly, an artificial gene is synthesized which comprises the light chain sequence (variable region sequence and constant region sequence) and the heavy chain sequence (variable region sequence and constant region sequence) of the anti-PD-L1 antibody of the present invention. The resultant gene is inserted into a vector (e.g., plasmid), which is then introduced into a host cell (e.g., mammal cell such as CHO cell). The host cell is cultured, and the antibody of interest is collected from the resultant culture. In the synthesis of the artificial gene, codons of the nucleotide sequence may be optimized.

The present invention provides a DNA encoding an anti-PD-L1 antibody comprising: (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3). CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5). The present invention also provides a DNA encoding a light chain of an anti-PD-L1 antibody comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) (the DNA of (a′)). Further, the present invention provides a DNA encoding a heavy chain of an anti-PD-L1 antibody comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5) (the DNA of (b)).

For (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5), reference should be had to the foregoing description. A DNA comprising the DNA of (a′) and the DNA of (′b) may be synthesized on commercial synthesizer. Restriction enzyme recognition sites, KOZAK sequences, poly-A addition signal sequences, promoter sequences, intron sequences or the like may be added to this DNA.

The present invention also provides a vector comprising the above-mentioned DNA encoding an anti-PD-L1 antibody.

As the vector, Escherichia coli-derived plasmids (e.g., pBR322, pBR325, pUC12 or pUC13); Bacillus subtilis-derived plasmids (e.g., pUB110, pTP5 or pC194), yeast-derived plasmids (e.g., pSH19 or pSH15); bacteriophages such as λ phage; animal viruses such as retrovirus or vaccinia virus; or insect pathogen viruses such as baculovirus may be used. In the Examples described later, pDC6 (Japanese Patent No. 5704753, U.S. Pat. No. 9,096,878, EU Patent 2385115, Hong Kong (China) patent HK1163739 and Australia Patent 2009331326) is used.

The vector may also comprise promoters, enhancers, splicing signals, poly-A addition signals, intron sequences, selection markers, SV40 replication origins, and so forth.

The present invention also provides a host cell transformed by the above vector. It is possible to prepare the anti-PD-L1 antibody of the invention by culturing the host cell and collecting the antibody of interest from the resultant culture. Therefore, the present invention also provides a method of preparing an antibody, comprising culturing the above-described host cell and collecting the anti-PD-L1 antibody of the invention from the culture. In the method of the present invention for preparing an antibody, a vector incorporating a DNA comprising a DNA encoding the light chain and a DNA encoding the heavy chain may be transfected into a host cell. Alternatively, a vector incorporating a DNA encoding the light chain and a vector incorporating a DNA encoding the heavy chain may be co-transfected into a host cell.

Examples of the host cell include, but are not limited to, bacterial cells (such as Escherichia bacteria, Bacillus bacteria or Bacillus subtilis), fungal cells (such as yeast or Aspergillus), insect cells (such as S2 cells or Sf cells), animal cells (such as CHO cells, COS cells, HeLa cells, C127 cells, 3T3 cells, BHK cells or HEK 293 cells) and plant cells. Among these, CHO-DG44 cell (CHO-DG44(dfhr⁻/⁻)) which is a dihydrofolate reductase deficient cell is preferable.

Introduction of a recombinant vector into a host cell may be performed by the methods disclosed in Molecular Cloning 2nd Edition, J. Sambrook et al., Cold Spring Harbor Lab. Press, 1989 (e.g., the calcium phosphate method, the DEAE-dextran method, transfection, microinjection, lipofection, electroporation, transduction, scrape loading, the shotgun method, etc.) or by infection.

The resultant transformant may be cultured in a medium, followed by collection of the anti-PD-L1 antibody of the present invention from the culture. When the antibody is secreted into the medium, the medium may be recovered, followed by isolation and purification of the antibody from the medium. When the antibody is produced within the transformed cells, the cells may be lysed, followed by isolation and purification of the antibody from the cell lysate.

Examples of the medium include, but are not limited to, OptiCHO medium, Dynamis medium, CD CHO medium, ActiCHO medium, FortiCHO medium, Ex-Cell CD CHO medium, BalanCD CHO medium, ProCHO 5 medium and Cellvento CHO-100 medium.

The pH of the medium varies depending on the cell to be cultured. Generally, a pH range from 6.8 to 7.6 is used; mostly, a pH range from 7.0 to 7.4 is appropriate.

When the cell to be cultured is CHO cells, culture may be performed by methods known to those skilled in the art. For example, it is usually possible to perform culturing in a gas-phase atmosphere having a CO₂ concentration of 0-40%, preferably 2-10%, at 30-39° C., preferably around 37° C.

The appropriate period of culture is usually from one day to three months, preferably from one day to three weeks.

Isolation and purification of the antibody may be performed by known methods. Known isolation/purification methods which may be used in the present invention include, but are not limited to, methods using difference in solubility (such as salting-out or solvent precipitation); methods using difference in molecular weight (such as dialysis, ultrafiltration, gel filtration or SDS-polyacrylamide gel electrophoresis); methods using difference in electric charge (such as ion exchange chromatography); methods using specific affinity (such as affinity chromatography), methods using difference in hydrophobicity (such as reversed phase high performance liquid chromatography); and methods using difference in isoelectric point (such as isoelectric focusing).

It is also possible to prepare the anti-PD-L1 antibody of the present invention by culturing a hybridoma which may be prepared by the method disclosed in the literature (Ikebuchi R, Konnai S, Okagavva T, Yokoyama K, Nakajima C, Suzuki Y, Murata S, Ohashi K, Immunology. 2014 August: 142(4):551-61). A hybridoma producing anti-PD-L1 antibody 6C11-3A11 is stored at the laboratory of the present inventor's (Laboratory of Infectious Diseases, Department of Disease Control, Faculty of Veterinary Medicine, Hokkaido University).

The PD-L1 antibody of the present invention may be used for detecting PD-L1. Therefore, the present invention provides a composition for detecting PD-L1, comprising the PD-L1 antibody as an active ingredient.

Detection of PD-L1 may be performed by such methods including, but are not limited to, immunohistochemical staining, immunocytochemical staining, flow cytometry, enzyme linked immunosorbent assay (ELISA) and Western blotting.

Analytes for detection may be exemplified by samples such as tissues or body fluids taken from organisms (e.g., blood (whole blood, plasma, serum, or specific cell such as erythrocyte, leukocyte or lymphocyte), urine, saliva, etc.); cell culture; and cultured cells (established cell lines, primary cultured cells, subcultured cells, etc.). The source of such analytes is not particularly limited. Examples include rat, canine, ovine, goat, porcine, feline, human, equine, bovine, water buffalo, yak, rabbit, mouse, hamster, and guinea pig.

The PD-L1 antibody of the present invention may be labeled with radioisotopes, enzymes, luminescent substances, fluorescent substances, biotin, or the like. If reaction with a primary antibody (the anti-PD-L1 antibody of the present invention) which specifically binds to a target molecule (PD-L1) is followed by reaction with a secondary antibody which binds to the primary antibody so as to detect the target molecule, it is suitable to label the secondary antibody.

Since PD-L1 is strongly expressed in cancer cells and virus-infected cells, the composition of the present invention may be used for diagnosis of cancers and/or infections. Usually, the amount (concentration) of PD-L1 in an analyte is determined based on the amount (concentration) of the complex of PD-L1 and anti-PD-L1 antibody. When the amount (concentration) of PD-L1 in the analyte is high compared to negative control (e.g., healthy surrounding tissue (connecting tissue, blood vessels, etc.)), the analyte may be diagnosed as suffering cancer and/or infection. Alternatively, if PD-L1 is detected in the analyte, the analyte may be diagnosed as suffering cancer and/or infection.

Examples of cancers and/or infections include, but are not limited to, neoplastic diseases (e.g., malignant melanoma, lung cancer, gastric cancer, renal cancer, breast cancer, bladder cancer, esophageal cancer, ovarian cancer and the like), leukemia, Johne's disease, anaplasmosis, bacterial mastitis, mycotic mastitis, mycoplasma infections (such as mycoplasma mastitis, mycoplasma pneumonia or the like), tuberculosis, Theileria orientalis infection, cryptosporidiosis, coccidiosis, trypanosomiasis and leishmaniasis.

The composition of the present invention can be used to select subject animals suitable for therapy using an anti-PD-L1 antibody. For example, animals satisfying the following two points may be considered as candidate animals.

1. A case diagnosed as suffering cancer (such as melanoma) or infection in pathological examination

2. A case found positive for anti-PD-L1 antibody

Negative control may be healthy surrounding tissue (connecting tissue, blood vessels, etc.), and positive control may be a case of cancer (such as melanoma) or infection. Basically, animals with a tumor which is positive in immunohistochemical staining of almost all regions may be subjected to clinical trial.

Subject animals are not particularly limited and may include rat, canine, ovine, goat, porcine, feline, human, equine, bovine, water buffalo, yak, rabbit, mouse, hamster, and guinea pig.

The composition of the present invention may further comprise reagents for detecting labels, diluents, lavage fluids, written instructions describing criteria for diagnosis/selection, and so on.

EXAMPLES

Hereinbelow, the present invention will be described in more detail with reference to the following Examples. However, the present invention is not limited to these Examples.

Example 1

1. Introduction

Programmed cell death 1 (PD-1), an immunoinhibitory receptor, and its ligand programmed cell death ligand 1 (PD-L1) are molecules identified by Prof. Tasuku Honjo et al., Kyoto University, as factors which inhibit excessive immune response and are deeply involved in immunotolerance. Recently, it has been elucidated that these molecules are also involved in immunosuppression in infections and tumors in various animals. In the subject Example, an anti-bovine PD-L1 monoclonal antibody was prepared by immunizing rats, and then a clone (6C11-3A11) capable of detecting canine PD-L1 was selected. Further, the present inventors performed immunohistochemical staining to examine whether or not this anti-bovine PD-L1 antibody 6C11-3A11 would be useful for detecting PD-L1 in canine malignant tumors (such as melanoma) and porcine/ovine infections.

2. Materials and Methods

2.1 Rat Anti-Bovine PD-L1 Monoclonal Antibody Producing Cells

The nucleotide sequence of bovine PD-L1 was identified (Ikebuchi R, Konnai S, Shirai T, Sunden Y, Murata S, Onuma M, Ohashi K, Vet Res. 2011 September 26:42:103). Based on the sequence information, a recombinant bovine PD-L1 was prepared. Rat was immunized in the footpad with this recombinant protein, and hybridomas were established by the iliac lymph node method. As a result, a plurality of hybridomas producing rat anti-bovine PD-L1 monoclonal antibodies were obtained (Ikebuchi R, Konnai S, Okagawa T, Yokoyama K, Nakajima C, Suzuki Y, Murata S, Ohashi K, Immunology 2014 August; 142(4):551-561). Rat anti-bovine PD-L1 antibody 6C11-3A11 is one of the monoclonal antibodies established from the above-described immunized rat.

2.2 Identification of Full-Length Canine PD-L1 Gene

To determine the full length of canine PD-L1 cDNA, PCR primers were first designed based on the putative nucleotide sequence of canine PD-L1 already registered at The National Center for Biotechnology Information (NCBI) (GenBank accession number; XM_541302). Briefly, primers to amplify the inner sequence of the open reading frame (ORF) of this gene were designed (cPD-1 inner F and R), and PCR was performed. For the amplified products, nucleotide sequences were determined with a capillary sequencer according to conventional methods. Further, to determine the nucleotide sequence of full-length PD-L1 cDNA, primers (cPD-L1 5′ GSP and 3′ GSP) were designed based on the canine PD-L1 cDNA sequences determined above. 5′-RACE and 3′-RACE were then performed using, respectively, the 5′-RACE system for rapid amplification of cDNA ends and 3′-RACE system for rapid amplification of cDNA ends (Invitrogen). The resultant gene fragments of interest were sequenced as described above (Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagavva Y, Nakajima C, Suzuki Y, Murata S, Ohashi K. PLoS One. 2014 June 10;9(6):e98415).

Primer (cPD-L1 inner F): ATGAGAATGTTTAGTGTCTT (SEQ ID NO: 22) Primer (cPD-L1 inner R): TTATGTCTCTTCAAATTGTATATC (SEQ ID NO: 23) Primer (cPD-L1 5′GSP): TTTTAGACAGAAAGTGA (SEQ ID NO: 24) Primer (cPD-L1 3′GSP): GACCAGCTCTTCTTGGGGAA (SEQ ID NO: 25) 2.3 Preparation of Canine PD-L1 Expressing COS-7 Cells

For preparing a canine PD-L1-EGFP expression plasmid, PCR was performed using a synthesized beagle PBMC-derived cDNA as a template and primers designed by adding BglII and EcoRI recognition sites on the 5′ side (cPD-L1-EGFP F and R). The resultant PCR products were digested with BglII (New England Biolabs) and EcoRI (Takara), and then purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics), followed by cloning into pEGFP-N2 vector (Clontech) similarly treated with restriction enzymes. The resultant expression plasmid of interest was extracted with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pEGFP-N2-cPD-L1.

Primer (cPD-L1-EGFP F): GAAGATCTATGAGAATGTTTAGTGTC (SEQ ID NO: 26) Primer (cPD-L1-EGFP R): GGAATTCTGTCTCTTCAAATTGTATATC (SEQ ID NO: 27)

COS-7 cells were subcultured at a density of 5×10⁴ cells/cm² in 6-well plates, and then cultured overnight in RPMI 1640 medium containing 10% inactivated fetal bovine serum and 0.01% L-glutamine at 37° C. in the presence of 5% CO₂. The pEGFP-N2-cPD-L1 or pEGFP-N2 (negative control) was introduced into COS-7 cells at 0.4 μg/cm² using Lipofectamine 2000 (Invitrogen). The cells were cultured for 48 hours (canine cPD-L1-EGFP expressing cell and EGFP expressing cell). In order to confirm the expression of PD-L1 in the thus prepared expressing cells, intracellular localization of enhanced green fluorescent protein (EGFP) was visualized with an inverted confocal laser microscope LSM700 (ZEISS) (Maekawa N, Konnai S, Ikebuchi R, Okagawa T, Adachi M, Takagi S, Kagawa Y, Nakajima C, Suzuki Y, Murata S, Ohashi K. PLoS One. 2014 June 10;9(6):e98415).

2.4 Cross-Reactivity of Rat Anti-Bovine PD-L1 Antibody 6C11-3A11 with Canine PD-L1

In order to confirm that rat anti-bovine PD-L1 antibody 6C11-3A11 specifically binds to canine PD-L1, flow cytometry was performed using the canine cPD-L1-EGFP expressing cell or the EGFP expressing cell prepared in 2.3 above. To 2×10⁵−1×10⁶ cells, 10 μg/ml of anti-bovine PD-L1 antibody 6C11-3A11 was added and reacted for 30 min at room temperature. After washing, the antibody binding to cell surfaces was detected with Altophycocyanine-labeled goat anti-rat Ig antibody (Beckman Coulter). For the analysis, FACS Verse (Becton, Dickinson and Company) was used. As a negative control antibody, rat IgG2a (κ) isotype control (BD Bioscience) was used. For every washing operation and dilution of antibodies, 10% inactivated goat serum-supplemented PBS was used. The results are shown in FIG. 1 .

2.5 CDR Analysis of Rat Anti-Bovine PD-L1 Antibody 6C11-3A11

The heavy chain and the light chain genes of rat anti-bovine PD-L1 antibody 6C11-3A11 were identified from a hybridoma producing the antibody by RACE method. The complementarity-determining regions (CDRs) of rat anti-bovine PD-L1 antibody 6C11-3A11 were determined using NCBI IGBLAST (www.ncbi.nlm.nih.gov/igblast/). The results are shown in FIG. 2 .

2.6 Immunohistochemical Staining of Canine Tumor Tissues and Ovine/Porcine Infected Tissues

In order to confirm that rat anti-bovine PD-L1 antibody 6C11-3A11 is applicable to PD-L1 immunohistochemical staining of canine tumors, formalin-fixed and paraffin-embedded canine tumor samples were immunohistochemically stained. According to conventional methods, the resultant samples were deparaffinized and then subjected to microwave treatment (5 min, twice) in citrate buffer. Subsequently, the samples were reacted with PD-L1 antibody 6C11-3A11 (400-fold dilution) for 30 min and then with Simple Stain Mouse MAX-PO (Rat) (Nichirei Bioscience) for 30 min. For coloring, diaminobenzidine (DAB) was reacted for 10 min.

The results are shown in FIGS. 3,4, 5-1, 5-2, 6 and 7 .

Anti-MelanA antibody, the only commercially available antibody specific to melanoma stained tumor cells very weakly (FIG. 3 , left). On the other hand, the PD-L1 antibody (6C11-3A11) established by the present inventors stained tumor cells very strongly (FIG. 3 , Right). The PD-L1 antibody (6C11-3A11) was capable of staining almost all cases of melanoma.

In canine melanoma tumor cells were found diffusely positive for the PD-L1 antibody (6C11-3A11). (Positive number/tested number=12/12; positive rate 100%)

In canine lymphoma, tumor cells were found diffusely positive for the PD-L1 antibody (6C11-3A11). In canine osteosarcoma, some tumor cells were stained intracellularly. In canine renal cell carcinoma, tumor cells were found diffusely positive in various tissue types.

In a case of ovine listeriosis, a PD-L1 staining image of a brain lesion of ovine listeriosis showing neurologic symptoms is shown in FIG. 6 , left panel. In an enlarged photograph of this image, expression of PD-L1 was observed in macrophages infiltrating into brain lesions (FIG. 6 , Right).

In a case of porcine circovirus type 2 infection, PD-L1 was stained with lymphoid follicles, and virus was stained in these cells (FIG. 7 , Left).

In a case of porcine mycoplasma pneumonia a great number of macrophages infiltrated pulmonary lesions, and PD-L1 was stained in these infiltrating cells (FIG. 7 , Right).

As described so far, anti-bovine PD-L1 antibody 6C11-3A11 may be used for detecting PD-L1 in various canine tumors (such as malignant melanoma) and ovine/porcine infections by immunohistochemical staining. This suggests the possibility of using anti-bovine PD-L1 antibody 6C11-3A11 for diagnosis in a multiple-animal and a multiple-disease manner.

Example 2

1. Introduction

Monoclonal antibodies may be produced by culturing hybridomas and purifying antibodies from the resultant culture supernatants. Alternatively, when the nucleotide sequence of an antibody of interest has been identified, a cell expressing the antibody may be prepared by transfecting cultured cells with a vector for expressing the nucleotide sequence; the thus prepared antibody expressing cell may be used as a substitute for hybridoma. In the subject Example, a method will be illustrated below in which an antibody is produced by a protein expression system using an expression vector and a mammalian cell.

2. Materials and Methods

2.1 Preparation of Rat Anti-Bovine PD-L1 Antibody 6C11-3A11 Expression Vector

Based on the nucleotide sequence of rat anti-bovine PD-L1 antibody 6C11-3A11 identified in 2.5 of Example 1 above, gene synthesis is performed so that NotI restriction enzyme recognition site, KOZAK sequence, antibody's light chain sequence, poly-A addition signal sequence (PABGH), promoter sequence (PCMV), SacI restriction enzyme recognition site, intron sequence (INRBG), KOZAK sequence, antibody's heavy chain sequence and XbaI restriction enzyme recognition site will be located in this order. In this case, codons of the antibody gene may have been optimized in advance depending on the type of the cell that is to express the gene. The synthesized gene strand is incorporated into an expression vector pDC6 (kindly provided by Prof. S. Suzuki, Research Center for Zoonosis Control. Hokkaido University) at the cloning site (NotI and XbaI restriction enzyme recognition sequences located downstream of PCMV and between INRBG and PABGH) using restriction enzyme recognition sequences so that the above-listed sequences will be located in the above-mentioned order to thereby construct a rat anti-bovine PD-L1 antibody expressing vector pDC6.

2.2 Expression of Rat Anti-Bovine PD-L1 Antibody 6C11-3A11

The rat anti-bovine PD-L1 antibody expressing vector pDC6 as prepared in 2.1 above is transfected into CHO-DG44 cells (CHO-DG44(dfhr⁻/⁻)) which are dihydrofolate reductase deficient cells, and high expression clones are selected by dot blotting. For increased expression, gene amplification treatment may be performed by adding load on cells in a medium containing 60 nM, 250 nM or 1000 nM methotrexate (Mtx). The thus prepared cells stably expressing rat anti-bovine PD-L1 antibody 6C11-3A11 are transferred to Mtx-free Opti-CHO medium. By culturing those cells under shaking for 14 days (125 rpm, 37° C., 5% CO₂), a culture supernatant containing the antibody of interest can be obtained. The antibody in the culture supernatant may be purified by known methods such as affinity chromatography or ion exchange chromatography for use in various experiments.

Example 3

1. Introduction

For the purpose of establishing a novel diagnosis method for tumor diseases, a rat-human chimeric anti-PD-L1 antibody is obtained in the subject Example by culturing Chinese hamster ovary cells (CHO cells) that will express a chimeric antibody gene in which the variable region gene of rat anti-bovine PD-L1 antibody 6C11-3A11 is combined with the constant region gene of human immunoglobulin (IgG4).

2. Materials and Methods

2.1 Preparation of Rat-Human Chimeric Anti-PD-L1 Expression Vector (FIG. 10 )

Hereinbelow, a rat-human chimeric anti-PD-L1 antibody is established using rat anti-bovine PD-L1 monoclonal antibody 6C11-3A11 as its variable region.

Briefly, heavy chain and light chain variable region genes were identified from a hybridoma producing the rat anti-bovine PD-L1 antibody 6C11-3A11. Further, a nucleotide sequence was prepared by linking the heavy chain and light chain variable region genes of the above rat antibody to the constant region of heavy chain IgG4 and the constant region of light chain Kappa of a known human antibody, respectively. After codon optimization, gene synthesis is performed so that NotI restriction enzyme recognition site, KOZAK sequence, chimeric antibody's light chain sequence, poly-A addition signal sequence (PABGH), promoter sequence (PCMV), SacI restriction enzyme recognition site, intron sequence (INRBG), KOZAK sequence, chimeric antibody's heavy chain sequence and XbaI restriction enzyme recognition site will be located in this order. The synthesized gene strand is incorporated into the expression vector pDC6 (kindly provided by Prof. S. Suzuki, Research Center for Zoonosis Control, Hokkaido University) at the cloning site (NotI and XbaI restriction enzyme recognition sequences located downstream of PCMV and between INRBG and PABGH) using restriction enzyme recognition sequences so that the above-listed sequences will be located in the above-mentioned order (FIG. 10 ). Thus, a rat-human chimeric anti-PD-L1 antibody expressing vector is constructed. This expression vector is transfected into CHO-DG44 cells (CHO-DG44(dfhr⁻/⁻)) which are dihydrofolate reductase deficient cells, and high expression clones are selected by dot blotting. For increased expression, gene amplification treatment may be performed by adding load on cells in a medium containing 60 nM, 250 nM or 1000 nM methotrexate (Mtx). The thus prepared cells stably expressing rat-human chimeric anti-PD-L1 antibody 6C11-3A11 are transferred to Mtx-free Opti-CHO medium. By culturing those cells under shaking for 14 days (125 rpm, 37° C., 5% CO₂), a culture supernatant containing the antibody of interest can be obtained. The antibody in the culture supernatant may be purified by known methods such as affinity chromatography or ion exchange chromatography for use in various experiments.

Example 4

1. Introduction

With respect to PD-L1 in canine tumors, a detection method by immunohistochemical staining with rat anti-bovine PD-L1 antibody 6G7-E1 was previously established, and the expression profiles in various tumors have been reported (Maekawa N, Konnai S, Okagawa T, Ikebuchi R, Izumi Y, Takagi S, Kagavva Y, Nakajima C, Suzuki Y, Kato Y, Murata S, Ohashi K. PLoS One. 2016 June 11(6):e0157176). In the subject Example, in order to examine whether rat anti-bovine PD-L1 antibody 6C11-3A11 is more useful than existing anti-PD-L1 antibody 6G7-E1 in expression analysis of PD-L1 in canine tumors, immunohistochemical staining of various canine tumors was performed to thereby directly compare the PD-L1 detection sensitivities of 6G7-E1 and 6C11-3A11.

2. Materials and Methods

2.1 Comparison by Flow Cytometry Using Canine PD-L1-EGFP Stably Expressing CHO-DG44 Cells (FIG. 11 )

First, in order to prepare canine PD-L1 membrane expressing cells, 2.5 μg of canine PD-L1-EGFP expression plasmid (pEGFP-N2-cPD-L1) prepared in 2.3 of Example 1 or pEGFP-N2 (negative control) was introduced into 4×10⁶ CHO-DG44 cells using Lipofectamine LTX (Invitrogen). Forty-eight hours later, the medium was exchanged with CD DG44 medium (Life Technologies) containing G418 (Enzo Life Science) 800 μg/ml, GlutaMAX supplement (Life Technologies) 20 ml/L, and 10% Pluronic F-68 (Life Technologies) 18 ml/L, followed by selection of stably expressing cells and cloning by limiting dilution. The thus prepared canine PD-L1 membrane expressing cell or EGFP expressing cell was reacted with rat anti-bovine PD-L1 antibody 6C11-3A11 or 6G7-E1 at room temperature for 30 min. After washing, antibodies binding to cell surfaces were detected with Allophycocyanine-labeled goat anti-rat Ig antibody (Beckman Coulter). For analysis, FACS Verse (Becton, Dickinson and Company) was used. As a negative control, rat IgG2a (κ) or IgM (κ) isotype control (BD Bioscience) was used. For every washing operation and dilution of antibodies, 10% inactivated goat serum-supplemented PBS was used.

The results are shown in FIG. 11 . Rat anti-bovine PD-L1 antibodies 6C11-3A11 and 6G7-E1 bound specifically to canine PD-L1 membrane expressing cells. The resultant fluorescence intensity was higher with 6C11-3A11 than with 6G7-E1, suggesting that 6C11-3A11 is an antibody with higher affinity.

2.2 Comparison of the Detection Sensitivities of Both Antibodies in PD-L1 Expression Analysis of Various Canine Tumors (Immunohistochemical Staining)

Using samples of canine skin squamous cell carcinoma (n=5), nasal adenocarcinoma (n=5), transitional cell carcinoma (n=5), anal sac gland carcinoma (n=5), soft tissue sarcoma (n=5) and osteosarcoma (n=5), immunohistochemical staining with rat anti-bovine PD-L1 antibody 6C11-3A11 was performed according to the method described in 2.6 of Example 1. With rat anti-bovine PD-L1 antibody 6G7-E1, immunohistochemical staining was performed in the same manner using sections derived from the same analytes. The final concentration of 6G7-E1 used on this occasion was 10 μg/ml, and biotin-labeled goal anti-rat IgM antibody (Jackson ImmunoResearch Laboratories) was used as a secondary antibody.

The results are shown in FIGS. 12 and 13 . While specific signals were not observed in squamous cell carcinoma, nasal adenocarcinoma, transitional cell carcinoma, anal sac gland carcinoma and soft tissue sarcoma upon staining with 6G7-E1, satisfactory positive reactions were obtained upon staining with 6C11-3A11. On the other hand, specific signals were also obtained with 6G7-E1 in osteosarcoma but staining with 6C11-3A11 provided higher signal intensities. The PD-L1 positive rate of these tumors obtained by 6C11-3A11 staining was 100% (5 out of 5 cases) in all of the above-listed tumor species excepting soft tissue sarcoma which turned out to be PD-L1 positive at a rate of 80% (4 out of 5 cases).

Subsequently, samples of oral malignant melanoma (n=17), mammary adenocarcinoma (n=10), histiocytic sarcoma (n=10), diffuse large B-cell lymphoma (n=10) and transmissible venereal tumor (n=4) were immunohistochemically stained with 6C11-3A11 to analyze PD-L1 expression therein.

The results are shown in FIG. 14 . The PD-L1 positive rate was 100% in oral malignant melanoma (17 out of 17 cases), 100% in mammary adenocarcinoma (10 out of 10 cases), 20% in histiocytic sarcoma (2 out of 10 cases), 20% in diffuse large B-cell lymphoma (2 out of 10 cases) and 0% in transmissible venereal tumor (0 out of 4 cases).

The above-described results revealed that 6C11-3A11 is superior to the existing anti-PD-L1 antibody 6G7-E1 in the detection of canine PD-L1.

Example 5

1. Introduction

Johne's disease is a bovine chronic infection caused by Mycobacterium avium subsp. paratuberculosis. In cattle affected with Johne's disease, PD-L1 expression has been confirmed in M. avium subsp. paratuberculosis-infected cells in ileal lesions which are a localized site of infection with this bacterium (Okagawa T, Konnai S, Nishimori A, Ikebuchi R, Mizorogi S, Nagata R, Kawaji S, Tanaka S, Kagawa Y, Murata S, Mori Y and Ohashi K. Infect Immun, 84:77-89, 2016). In the subject Example, immunohistochemical staining of ileal lesions of cattle with Johne's disease was performed in order to examine whether rat anti-bovine PD-L1 antibody 6C11-3A11 could be used for detecting bovine PD-L1 or not.

2. Materials and Methods

2.1. Construction of Bovine PD-L1 Expressing Cells

The nucleotide sequence of the full-length cDNA of bovine PD-L1 gene (GenBank accession number AB510902; Ikebuchi R, Konnai S, Shirai T, Sunden Y, Murata S, Onuma M, Ohashi K, Vet. Res, 2011 September 26;42:103) was determined. Based on the resultant nucleotide sequence, bovine PD-L1 membrane expressing cells were prepared. First, for preparing a bovine PD-L1 expressing plasmid, PCR was performed using a synthesized bovine PBMC-derived cDNA as a template and primers having NheI and XhoI recognition sites added on the 5′ side (boPD-L1-EGFP F and R). The PCR products were digested with NheI (Takara) and XhoI (Takara), purified with FastGene Gel/PCR Extraction Kit (NIPPON Genetics) and cloned into pEGFP-N2 vector (Clontech) that had been subjected to similar restriction enzyme treatments. The resultant expression plasmid of interest was extracted with QIAGEN Plasmid Midi kit (Qiagen) and stored at −30° C. until use in experiments. Hereinafter, the thus prepared expression plasmid is designated as pEGFP-N2-boPD-L1.

Primer (boPD-L1-EGFP F): (SEQ ID NO: 124) CTAGCTAGCACCATGAGGATATATAGTGTCTTAAC Primer (boPD-L1-EGFP R): (SEQ ID NO: 125) CAATCTCGAGTTACAGACAGAAGATGACTGC

Bovine PD-L1 membrane expressing cells were prepared by the procedures described below. First, 2.5 μg of pEGFP-N2-boPD-L1 or pEGFP-N2 (negative control) was introduced into 4×10⁶ CHO-DG44 cells using Lipofectannne LTX (Invitrogen). Forty-eight hours later, the medium was exchanged with CD DG44 medium (Life Technologies) containing G418 (Enzo Life Science) 800 μg/ml, GlutaMAX supplement (Life Technologies) 20 ml/L, and 10% Pluronic F-68 (Life Technologies) 18 ml/L; thereafter, selection was performed simultaneously with cloning by limiting dilution (bovine PD-L1 expressing cell and EGFP expressing cell). In order to confirm the expression of bovine PD-L1 in the thus prepared bovine PD-L1 expressing cell, intracellular localization of EGFP was visualized with an inverted confocal laser microscope LSM700 (ZEISS).

2.2. Binding Specificity of Rat Anti-Bovine PD-L1 Antibody 6C11-3A11 to Bovine PD-L1

It was confirmed by flow cytometry that rat anti-bovine PD-L1 antibody 6C11-3A11 specifically binds to the bovine PD-L1 expressing cell (described above). First, rat anti-bovine PD-L1 antibody 6C11-3A11 or rat IgG2a (κ) isotype control (BD Biosciences) as a negative control was reacted with the bovine PD-L1 expressing cell or the EGFP expressing cell (negative control) at room temperature for 30 min. After washing, APC-labeled anti-rat Ig goat antibody (Southern Biotech) was reacted at room temperature for 30 min. After washing, antibodies bound to cell surfaces were detected by FACS Verse (BD Biosciences). For every washing operation and dilution of antibody, PBS supplemented with 1% bovine serum albumin (Sigma-Aldrich) was used.

The results are shown in FIG. 15 . It was revealed that rat anti-bovine PD-L1 antibody 6C11-3A11 binds specifically to the bovine PD-L1 expressing cell.

2.3. Immunohistochemical Staining Using Tissue Samples from Infected Cattle

In order to confirm that rat anti-bovine PD-L1 antibody 6C11-3A11 is applicable to PD-L1 immunohistochemical staining of bovine tissues, immunohistochemical staining was performed with formalin-fixed, paraffin-embedded bovine tissue samples. Briefly, ilium tissue blocks from cattle which naturally developed Johne's disease (#1, presenting clinical symptoms of Johne's disease such as diarrhea and severe emaciation), cattle experimentally infected with M. avium subsp. paratuberculosis (#65, clinical symptoms such as shedding of M. avium subsp. paratuberculosis and diarrhea were observed; Okagawa T, Konnai S, Nishimori A, Ikebuchi R, Mizorogi S, Nagata R, Kawaji S, Tanaka S, Kagavva Y, Murata S, Mori Y and Ohashi K. Infect Immun. 84:77-89, 2016) and uninfected control cattle (C #6) (the blocks kindly provided by Dr. Yasuyuki Mori, National Institute of Animal Health, National Agriculture and Food Research Organization) were used for staining. According to conventional methods, the stained samples were deparaffinized and then subjected to microwave treatment (5 min, twice) in citrate buffer. Subsequently, the samples were reacted with rat anti-bovine PD-L1 antibody 6C11-3A11 (400-fold dilution) for 30 min and then with Simple Stain Mouse MAX-PO (Rat) (Nichirei Bioscience) for 30 min. Finally, the samples were reacted with diaminobenzidine (DAB) for 10 min for coloring, followed by observation with an optical microscope.

The results are shown in FIG. 16 . Rat anti-bovine PD-L1 antibody 6C11-3A11 detected expression of PD-L1 in M. avium subsp. paratuberculosis-infected cells (confirmed by Ziehl-Neelsen staining) in ileal lesions of cattle #1 that naturally developed Johne's disease and experimentally infected cattle #65 (FIG. 16 a, b ). On the other hand, PD-L1 was not expressed in the ileum of uninfected cattle (C #6), so reaction of rat anti-bovine PD-L1 antibody 6C11-3A11 (non-specific reaction) was not recognized (FIG. 16 a ).

As described above, it was shown that rat anti-bovine PD-L1 antibody 6C11-3A11 can be used for detecting PD-L1 in bovine tissues by immunohistochemical staining.

All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

INDUSTRIAL APPLICABILITY

The anti-PD-L1 antibody of the present invention is applicable to diagnosis of cancers and/or infections. Further, the anti-PD-L1 antibody of the present invention is also applicable to selection of subject animals suitable for therapy with anti-PD-L1 antibodies.

SEQUENCE LISTING FREE TEXT <SEQ ID NO: 1>

<SEQ ID NO: 2>

<SEQ ID NO: 3> SEQ ID NO: 3 shows the amino acid sequence of CDR1 of the  heavy chain variable region (VH) of anti-PD-LI antibody  6C11-3A11 (IgG2a). GYTFIDYI <SEQ ID NO: 4>

<SEQ ID NO: 5>

<SEQ ID NO: 6>

<SEQ ID NO: 7>

<SEQ ID NO: 8> SEQ ID NO: 8 shows the amino acid sequence of the light chain (kappa chain) constant region of anti-PD-L1 antibody 6C11-3A11 (IgG2a). RADAAPTVSIFPPSTEQLATGGASVVCLMNNFYPRDISVKWKIDGTERRDGVLDSVTDQDS KDSTYSMSSTLSLTKADYESHNLYTCEVVHKTSSSPVVKSFNRNEC* <SEQ ID NO: 9> SEQ ID NO: 9 shows the amino acid sequence of the heavy chain constant region (CH) of anti-PD-L1 antibody 6C11-3A11  IgG2a). AETTAPSVYPLAPGTALKSNSMVTLGCLVKGYFPEPVTVTWNSGALSSGVHTFPAVLQSGL YTLTSSVTVPSSTWSSQAVTCNVAHPASSTKVDKKIVPRECNPCGCTGSEVSSVFIFPPKT KDVLTITLTPKVTCVVVDISQNDPEVRFSWFIDDVEVHTAQTHAPEKQSNSTLRSVSELPI VHRDWLNGKTFKCKVNSGAFPAPIEKSISKPEGTPRGPQVYTMAPPKEEMTQSQVSITCMV KGFYPPDIYTEWKMNGQPQENYKNTPPTMDTDGSYFLYSKLNVKKETWQQGNTFTCSVLHE GLHNHHTEKSLSHSPGK* <SEQ ID NO: 10> SEQ ID NO: 10 shows the amino acid sequence (GenBank: #V01241.1) of the light chain (kappa chain) constant region  of a rat antibody (IgG2a). ADAAPTVSIFPPSTEQLATGGASVVCLMNNFYPRDISVKWKIDGTERRDGVLDSVTDQDSK DSTYSMSSTLSLTKADYESHNLYTCEVVHKTSSPVVKSFNRNEC* <SEQ ID NO: 11> SEQ ID NO: 11 shows the amino acid sequence (GenBank: #X16129.1) of the light chain(kappa chain) constant region of a rat antibody (IgG2a). RADAAPTVSIFPPSTEQLATGGASVVCLMNNFYPRDISVKWKIDGTERRDGVLDSVTDQDS KDSTYSMSSTLSLSKADYESHNLYTCEVVHKTSSSPVVKSFNRNEC <SEQ ID NO: 12> SEQ ID NO: 12 shows the amino acid sequence (GenBank:  #DQ402417.1) of the light chain (kappa chain) constant region of a rat antibody (IgG2a). AAPTVSIFPPSMEQLTSGGATVVCFVNNFYPRDISVKWKIDGSEQRDGVLDSVTDQDSKDS TYSMSSTLSLTKVEYERHNLYTCEVVHKTSSSPVVKSFNRNEC* <SEQ ID NO: 13> SEQ ID NO: 13 shows the amino acid sequence (GenBank:  #DQ402472.1) of the CH of a rat antibody (IgG2a). APSVYPLAPGTALKSNSMVTLGCLVKGYFPEPVTVTWNSGALSSGVHTFPAVLQSGLYTLT SSVTVPSSTWSSQAVTCNVAHPASSTKVDKKIVPRECNPCGCTGSEVSSVFIFPPKTKDVL TITLTPKVTCVVVDISQNDPEVRFSWFIDDVEVHTAQTHAPEKQSNSTLRSVSELPIHRDW LNGKTFKCKVNSGAFPAPIEKSISKPEGTPRGPQVYTMAPPKEEMTQSQVSITCMVKGFYP PDIYTEWKMNGQPQENYKNTPPTMDTDGSYFLYSKLNVKKETWQQGNTFTCSVLHEGLHNH HTEKSLSHSPGK* <SEQ ID NO: 14> SEQ ID NO: 14 shows the nucleotide sequence of the VL of  anti-PD-L1 antibody 6C11-3A11(IgG2a). ATGAGGGTCCAGATTCAGTTTTGGGGGCTTCTTCTGCTCTGGACATCAGGTATACAGTGTG ATGTCCAGATGACCCAGTCTCCATCTAATCTTGCTGCCTCTCCTGGAGAAAGTGTTTCCAT CAATTGCAAGGCAAGTAAGAGCATTAGCAAGTATTTAGCCTGGTATCAACAGAAACCTGGG AAAGCAAATAAGCTTCTTATCTACTCTGGGTCAACTTTGCAATCTGGAACTCCATCGAGGT TCAGTGGCAGTGGATCTGGTACAGATTTCACTCTCACCATCAGAAACCTGGAGCCTGAAGA TTTTGGACTCTATTACTGTCAACAGCATAATGAATACCCGCTCACGTTCGGTTCTGGGACC AAGCTGGAGATCAAA <SEQ ID NO: 15> SEQ ID NO: 15 shows the nucleotide sequence of the VH of  anti-PD-L1 antibody 6C11-3A11 (IgG2a). ATGGGATGGATCTGTATCATCTTTCTTGTGGCAATAGCTACAGGTGCCCACTCCCAGGTCA AGCTGCTGCAGTCTGGGGCTGCACTGGTGAAGCCTGGGGACTCTGTGAAGATGTCTTGCAA AGCTTCTGGTTATACATTCACTGACTACATTATACACTGGGTGAAGCAGAGTCATGGAAAA AGCCTTGAGTGGATTGGTTATATTAATCCTGACAGTGGTGGTAATAACTACAATGAAAAGT TCAAGAGCAAGGCCACATTGACTGTAGACAAATCCAGCAGCACAGCCTATATGGAGTTTAG CAGATTGACATCTGAGGATTCTGCAATCTACTACTGTGCAAGAGGGATTACCATGATGGTA GTTATTAGCCACTGGAAGTTTGACTTCTGGGGCCCAGGAACCATGGTCACCGTGTCCTCA <SEQ ID NO: 16> SEQ ID NO: 16 shows the nucleotide sequence of the light  chain (kappa chain) constant region of anti-PD-L1 antibody   6C11-3A11 (IgG2a). CGGGCTGATGCTGCACCAACTGTATCTATCTTCCCACCATCCACGGAACAGTTAGCAACTG GAGGTGCCTCAGTCGTGTGCCTCATGAACAACTTCTATCCCAGAGACATCAGTGTCAAGTG GAAGATTGATGGCACTGAACGACGAGATGGTGTCCTGGACAGTGTTACTGATCAGGACAGC AAAGACAGCACGTACAGCATGAGCAGCACCCTCTCGTTGACCAAGGCTGACTATGAAAGTC ATAACCTCTATACCTGTGAGGTTGTTCATAAGACATCATCCTCACCCGTCGTCAAGAGCTT CAACAGGAATGAGTGTTAG <SEQ ID NO: 17> SEQ ID NO: 17 shows the nucleotide sequence of the CH of  anti-PD-L1 antibody 6C11-3A11 (IgG2a). GCTGAAACAACAGCCCCATCTGTCTATCCACTGGCTCCTGGAACTGCTCTCAAAAGTAACT CCATGGTGACCCTGGGATGCCTGGTCAAGGGCTATTTCCCTGAGCCAGTCACCGTGACCTG GAACTCTGGAGCCCTGTCCAGCGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGGACTC TACACTCTCACCAGCTCAGTGACTGTACCCTCCAGCACCTGGTCCAGCCAGGCCGTCACCT GCAACGTAGCCCACCCGGCCAGCAGCACCAAGGTGGACAAGAAAATTGTGCCAAGGGAATG CAATCCTTGTGGATGTACAGGCTCAGAAGTATCATCTGTCTTCATCTTCCCCCCAAAGACC AAAGATGTGCTCACCATCACTCTGACTCCTAAGGTCACGTGTGTTGTGGTAGACATTAGCC AGAATGATCCCGAGGTCCGGTTCAGCTGGTTTATAGATGACGTGGAAGTCCACACAGCTCA GACTCATGCCCCGGAGAAGCAGTCCAACAGCACTTTACGCTCAGTCAGTGAACTCCCCATC GTGCACCGGGACTGGCTCAATGGCAAGACGTTCAAATGCAAAGTCAACAGTGGAGCATTCC CTGCCCCCATCGAGAAAAGCATCTCCAAACCCGAAGGCACACCACGAGGTCCACAGGTATA CACCATGGCGCCTCCCAAGGAAGAGATGACCCAGAGTCAAGTCAGTATCACCTGCATGGTA AAAGGCTTCTATCCCCCAGACATTTATACGGAGTGGAAGATGAACGGGCAGCCACAGGAAA ACTACAAGAACACTCCACCTACGATGGACACAGATGGGAGTTACTTCCTCTACAGCAAGCT CAATGTAAAGAAAGAAACATGGCAGCAGGGAAACACTTTCACGTGTTCTGTGCTGCATGAG GGCCTGCACAACCACCATACTGAGAAGAGTCTCTCCCACTCTCCTGGTAAATGA <SEQ ID NO: 18> SEQ ID NO: 18 shows the nucleotide sequence (GenBank:  #V01241.1) of the light chain (kappa chain) constant region of a rat antibody (IgG2a). GGGCTGATGCTGCACCAACTGTATCTATCTTCCCACCATCCACGGAACAGTTAGCAACTGG AGGTGCCTCAGTCGTGTGCCTCATGAACAACTTCTATCCCAGAGACATCAGTGTCAAGTGG AAGATTGATGGCACTGAACGACGAGATGGTGTCCTGGACAGTGTTACTGATCAGGACAGCA AAGACAGCACGTACAGCATGAGCAGCACCCTCTCGTTGACCAAGGCTGACTATGAAAGTCA TAACCTCTATACCTGTGAGGTTGTTCATAAGACATCATCCTCACCCGTCGTCAAGAGCTTC AACAGGAATGAGTGTTAG <SEQ ID NO: 19> SEQ ID NO: 19 shows the nucleotide sequence (GenBank:  #X16129.1) of the light chain (kappa chain) constant region of a rat antibody (IgG2a). CGGGCTGATGCTGCACCAACTGTATCTATCTTCCCACCATCCACGGAACAGTTAGCAACTG GAGGTGCCTCAGTCGTGTCCTCATGAACAACTTCTATCCCAGAGACATCAGTGTCAAGTGG AAGATTGATGGCACTGAACGACGAGATGGTGTCCTGGACAGTGTTACTGATCAGGACAGCA AAGACAGCACGTACAGCATGAGCAGCACCCTCTCGTTGTCCAAGGCTGACTATGAAAGTCA TAACCTCTATACCTGTGAGGTTGTTCATAAGACATCATCCTCACCCGTCGTCAAGAGCTTC AACAGGAATGAGTGTTAG <SEQ ID NO: 20> SEQ ID NO: 20 shows the nucleotide sequence (GenBank:  #DQ402471.1) of the light chain (kappa chain) constant region of a rat antibody (IgG2a). GCCGCACCAACTGTATCCATCTTCCCACCATCCATGGAACAGTTAACATCTGGAGGTGCCA CAGTCGTGTGCTTCGTGAACAACTTCTATCCCAGAGACATCAGTGTCAAGTGGAAGATTGA TGGCAGTGAACAACGAGATGGTGTCCTGGACAGTGTTACTGATCAGGACAGCAAAGACAGC ACGTACAGCATGAGCAGCACCCTCTCGTTGACCAAGGTTGAATATGAAAGGCATAACCTCT ATACCTGTGAGGTTGTTCATAAGACATCATCCTCACCCGTCGTCAAGAGCTTCAACAGGAA TGAGTGTTAG <SEQ ID NO: 21> SEQ ID NO: 21 shows the nucleotide sequence (GenBank:  #DQ402472.1) of the CH of a rat antibody (IgG2a). CAGCCCCCTCTGTCTATCCACTGGCTCCTGGAACTGCTCTCAAAAGTAACTCCATGGTGAC CCTGGGATGCCTGGTCAAGGGCTATTTCCCTGAGCCAGTCACCGTGACCTGGAACTCTGGA GCCCTGTCCAGCGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGGACTCTACACTCTCA CCAGCTCAGTGACTGTACCCTCCAGCACCTGGTCCAGCCAGGCCGTCACCTGCAACGTAGC CCACCCGGCCAGCAGCACCAAGGTGGACAAGAAAATTGTGCCAAGGGAATGCAATCCTTGT GGATGTACAGGCTCAGAAGTATCATCTGTCTTCATCTTCCCCCCAAAGACCAAAGATGTGC TCACCATCACTCTGACTCCTAAGGTCACGTGTGTTGTGGTAGACATTAGCCAGAATGATCC CGAGGTCCGGTTCAGCTGGTTTATAGATGACGTGGAAGTCCACACAGCTCAGACTCATGCC CCGGAGAAGCAGTCCAACAGCACTTTACGCTCAGTCAGTGAACTCCCCATCGTGCACCGGG ACTGGCTCAATGGCAAGACGTTCAAATGCAAAGTCAACAGTGGAGCATTCCCTGCCCCCAT CGAGAAAAGCATCTCCAAACCCGAAGGCACACCACGAGGTCCACAGGTATACACCATGGCG CCTCCCAAGGAAGAGATGACCCAGAGTCAAGTCAGTATCACCTGCATGGTAAAAGGCTTCT ATCCCCCAGACATTTATACGGAGTGGAAGATGAACGGGCAGCCACAGGAAAACTACAAGAA CACTCCACCTACGATGGACACAGATGGGAGTTACTTCCTCTACAGCAAGCTCAATGTAAAG AAAGAAACATGGCAGCAGGGAAACACTTTCACGTGTTCTGTGCTGCATGAGGGCCTGCACA ACCACCATACTGAGAAGAGTCTCTCCCACTCTCCTGGTAAATGA <SEQ ID NOS: 22 to 27> SEQ ID NOS: 22 to 27 show the nucleotide sequences of primers cPD-L1 inner F, cPD-L1 inner R, cPD-L1 5′GSP,cPD-L1 3′GSP, cPD-L1-EGFP F and cPD-L1-EGFP R, in this order. <SEQ ID NO: 28> SEQ ID NO: 28 shows the amino acid sequence of the light  chain (kappa chain) constant region of a human antibody. <SEQ ID NO: 29> SEQ ID NO: 29 shows the nucleotide sequence of the light  chain (kappa chain) constant region of a human antibody. <SEQ ID NO: 30> SEQ ID NO: 30 shows the amino acid sequence of the CH (CH1—CH3) of a human antibody (IgG4 variant 1). <SEQ ID NO: 31> SEQ ID NO: 31 shows the nucleotide sequence of the CH  (CH1—CH3) of a human antibody (IgG4 variant 1). <SEQ ID NO: 32> SEQ ID NO: 32 shows the amino acid sequence of the CH  (CH1—CH3) of a human antibody (IgG4 variant 2). <SEQ ID NO: 33> SEQ ID NO: 33 shows the nucleotide sequence of the CH  (CH1—CH3) of a human antibody (IgG4 variant 2). <SEQ ID NO: 34> SEQ ID NO: 34 shows the amino acid sequence of the CH  (CH1—CH3) of a human antibody (IgG4 variant 3). <SEQ ID NO: 35> SEQ ID NO: 35 shows the nucleotide sequence of the CH  (CH1—CH3) of a human antibody (IgG4 variant 3). <SEQ ID NO: 36> SEQ ID NO: 36 shows the amino acid sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 37> SEQ ID NO: 37 shows the nucleotide sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 38> SEQ ID NO: 38 shows the amino acid sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 39> SEQ ID NO: 39 shows the nucleotide sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 40> SEQ ID NO: 40 shows the amino acid sequence of the light  chain (kappa chain)constant region of a mouse antibody. <SEQ ID NO: 41> SEQ ID NO: 41 shows the nucleotide sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 42> SEQ ID NO: 42 shows the amino acid sequence of the light  chain (kappa chain) constant region of a mouse antibody, <SEQ ID NO: 43> SEQ ID NO: 43 shows the nucleotide sequence of the light  chain (kappa chain) constant region of a mouse antibody. <SEQ ID NO: 44> SEQ ID NO: 44 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG1 variant 1). <SEQ ID NO: 45> SEQ ID NO: 45 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG1 variant 1). <SEQ ID NO: 46> SEQ ID NO: 46 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG1 variant 2). <SEQ ID NO: 47> SEQ ID NO: 47 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG1 variant 2). <SEQ ID NO: 48> SEQ ID NO: 48 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2a variant 1). <SEQ ID NO: 49> SEQ ID NO: 49 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2a variant 1). <SEQ ID NO: 50> SEQ ID NO: 50 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2a variant 2). <SEQ ID NO: 51> SEQ ID NO: 51 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2a variant 2). <SEQ ID NO: 52> SEQ ID NO: 52 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2b variant 1). <SEQ ID NO: 53> SEQ ID NO: 53 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2b variant 1). <SEQ ID NO: 54> SEQ ID NO: 54 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2b variant 2). <SEQ ID NO: 55> SEQ ID NO: 55 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2b variant 2). <SEQ ID NO: 56> SEQ ID NO: 56 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 1). <SEQ ID NO: 57> SEQ ID NO: 57 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 1). <SEQ ID NO: 58> SEQ ID NO: 58 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 2). <SEQ ID NO: 59> SEQ ID NO: 59 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 2). <SEQ ID NO: 60> SEQ ID NO: 60 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 3). <SEQ ID NO: 61> SEQ ID NO: 61 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG2c variant 3). <SEQ ID NO: 62> SEQ ID NO: 62 shows the amino acid sequence of the CH  (CH1—CH3) of a mouse antibody (IgG3). <SEQ ID NO: 63> SEQ ID NO: 63 shows the nucleotide sequence of the CH  (CH1—CH3) of a mouse antibody (IgG3). <SEQ ID NO: 64> SEQ ID NO: 64 shows the amino acid sequence of the light   chain (lambda chain) constant region of a bovine antibody. <SEQ ID NO: 65> SEQ ID NO: 65 shows the nucleotide sequence of the light  chain (lambda chain) constant region of a bovine antibody. <SEQ ID NO: 66> SEQ ID NO: 66 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG1 variant 1). <SEQ ID NO: 67> SEQ ID NO: 67 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG1 variant 1). <SEQ ID NO: 68> SEQ ID NO: 68 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG1 variant 2). <SEQ ID NO: 69> SEQ ID NO: 69 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG1 variant 2). <SEQ ID NO: 70> SEQ ID NO: 70 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG1 variant 3). <SEQ ID NO: 71> SEQ ID NO: 71 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG4 variant 3). <SEQ ID NO: 72> SEQ ID NO: 72 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG2 variant 1). <SEQ ID NO: 73> SEQ ID NO: 73 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG2 variant 1). <SEQ ID NO: 74> SEQ ID NO: 74 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG2 variant 2). <SEQ ID NO: 75> SEQ ID NO: 75 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG2 variant 2). <SEQ ID NO: 76> SEQ ID NO: 76 shows the amino acid sequence of the CH  (CH1-CH3) of a bovine antibody (IgG2 variant 3). <SEQ ID NO: 77> SEQ ID NO: 77 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG2 variant 3). <SEQ ID NO: 78> SEQ ID NO: 78 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG3 variant 1). <SEQ ID NO: 79> SEQ ID NO: 79 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG3 variant 1). <SEQ ID NO: 80> SEQ ID NO: 80 shows the amino acid sequence of the CH  (CH1—CH3) of a bovine antibody (IgG3 variant 2). <SEQ ID NO: 81> SEQ ID NO: 81 shows the nucleotide sequence of the CH  (CH1—CH3) of a bovine antibody (IgG3 variant 2). <SEQ ID NO: 82> SEQ ID NO: 82 shows the amino acid sequence of the light  chain (lambda chain) constant region of a canine antibody. <SEQ ID NO: 83> SEQ ID NO; 83 shows the nucleotide sequence of the light  chain (lambda chain) constant region of a canine antibody. <SEQ ID NO: 84> SEQ ID NO: 84 shows the amino acid sequence of the CH  (CH1—CH3) of a canine antibody (IgG-D). <SEQ ID NO: 85> SEQ ID NO: 85 shows the nucleotide sequence of the CH  (CH1—CH3) of a canine antibody (IgG-D). <SEQ ID NO: 86> SEQ ID NO: 86 shows the amino acid sequence of the light  chain (kappa chain) constant region of an ovine antibody. <SEQ ID NO: 87> SEQ ID NO: 87 shows the nucleotide sequence of the light  chain (kappa chain) constant region of an ovine antibody. <SEQ ID NO: 88> SEQ ID NO: 88 shows the amino acid sequence of the light  chain (lambda chain) constant region of an ovine antibody. <SEQ ID NO: 89> SEQ ID NO: 89 shows the nucleotide sequence of the light  chain (lambda chain) constant region of an ovine antibody. <SEQ ID NO: 90> SEQ ID NO: 90 shows the amino acid sequence of the CH  (CH1—CH3) of an ovine antibody (IgG1). <SEQ ID NO: 91> SEQ ID NO: 91 shows the nucleotide sequence of the CH  (CH1—CH3) of an ovine antibody (IgG1). <SEQ ID NO: 92> SEQ ID NO: 92 shows the amino acid sequence of the CH  (CH1—CH3) of an ovine antibody (IgG2). <SEQ ID NO: 93> SEQ ID NO: 93 shows the nucleotide sequence of the CH  (CH1—CH3) of an ovine antibody (IgG2). <SEQ ID NO: 94> SEQ ID NO: 94 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG1^(a)). <SEQ ID NO: 95> SEQ ID NO: 95 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG1^(a)). <SEQ ID NO: 96> SEQ ID NO: 96 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG1^(b)). <SEQ ID NO: 97> SEQ ID NO: 97 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG1^(b)). <SEQ ID NO: 98> SEQ ID NO: 98 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG2^(a)). <SEQ ID NO: 99> SEQ ID NO: 99 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG2^(a)). <SEQ ID NO: 100> SEQ ID NO: 100 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG2^(b)). <SEQ ID NO: 101> SEQ ID NO: 101 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG2^(b)). <SEQ ID NO: 102> SEQ ID NO: 102 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG3). <SEQ ID NO: 103> SEQ ID NO: 103 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG3). <SEQ ID NO: 104> SEQ ID NO: 104 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG4^(a)). <SEQ ID NO: 105> SEQ ID NO: 105 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG4^(a)). <SEQ ID NO: 106> SEQ ID NO: 106 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG4^(b)). <SEQ ID NO: 107> SEQ ID NO: 107 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG4^(b)). <SEQ ID NO: 108> SEQ ID NO: 108 shows the amino acid sequence of the CH  (CH1—CH3)of a porcine antibody (IgG5^(a)). <SEQ ID NO: 109> SEQ ID NO: 109 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG5^(a)). <SEQ ID NO: 110> SEQ ID NO: 110 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG5^(b)). <SEQ ID NO: 111> SEQ ID NO: 111 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG5b). <SEQ ID NO: 112> SEQ ID NO: 112 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG6^(a)). <SEQ ID NO: 113> SEQ ID NO: 113 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG6^(a)). <SEQ ID NO: 114> SEQ ID NO: 114 shows the amino acid sequence of the CH  (CH1—CH3) of a porcine antibody (IgG6^(b)). <SEQ ID NO: 115> SEQ ID NO: 115 shows the nucleotide sequence of the CH  (CH1—CH3) of a porcine antibody (IgG6^(b)). <SEQ ID NO: 116> SEQ ID NO: 116 shows the amino acid sequence of the light  chain (estimated to be Ig lambda) constant region (CL) of  a water buffalo antibody. <SEQ ID NO: 117> SEQ ID NO: 117 shows the nucleotide sequence of the light  chain (estimated to be Ig lambda) constant region (CL) of  a water buffalo antibody. <SEQ ID NO: 118> SEQ IlD NO: 118 shows the amino acid sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG1). <SEQ ID NO: 119> SEQ ID NO: 119 shows the nucleotide sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG1). <SEQ ID NO: 120> SEQ ID NO: 120 shows the amino acid sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG2). <SEQ ID NO: 121> SEQ ID NO: 121 shows the nucleotide sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG2). <SEQ ID NO: 122> SEQ ID NO: 122 shows the amino acid sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG3). <SEQ ID NO: 123> SEQ ID NO: 123 shows the nucleotide sequence of the CH  (CH1—CH3) of a water buffalo antibody (estimated to be  IgG3). <SEQ ID NO: 124> SEQ ID NO: 124 shows the nucleotide sequence of prime  boPD-L1-EGFP F. <SEQ ID NO: 125> SEQ ID NO: 125 shows the nucleotide sequence of primer  boPD-L1-EGFP R. 

The invention claimed is:
 1. A method to diagnose cancer and/or infection comprising contacting in vitro one or more cells from a subject with an antibody; and determining if said antibody binds to said one or more cells, wherein increased binding to said cells as compared to a control is indicative of cancer and/or infection, wherein the cancer is a cancer where cancer cells express PD-L1, wherein the infection is an infection where infected cells express PD-L1, and wherein said antibody is an anti-PD-L1 antibody comprising (a) a light chain comprising CDR1 having the amino acid sequence of KSISKY (SEQ ID NO: 1), CDR2 having the amino acid sequence of SGS and CDR3 having the amino acid sequence of QQHNEYPLT (SEQ ID NO: 2) and (b) a heavy chain comprising CDR1 having the amino acid sequence of GYTFTDYI (SEQ ID NO: 3), CDR2 having the amino acid sequence of INPDSGGN (SEQ ID NO: 4) and CDR3 having the amino acid sequence of ARGITMMVVISHWKFDF (SEQ ID NO: 5), wherein said antibody binds ovine, bovine, porcine or canine PD-L1 proteins, and wherein the cancer is not malignant melanoma.
 2. The method of claim 1, further comprising selecting the subject as a candidate for an anti-PD-L1 antibody therapy.
 3. The method of claim 1, further comprising administering an anti-PD-L1 antibody therapy to the subject.
 4. The method of claim 1, wherein the subject is selected from the group consisting of canine, ovine, porcine and bovine.
 5. The method of claim 1, wherein the antibody is derived from rat.
 6. The method of claim 5, wherein the antibody is a rat anti-bovine PD-L1 antibody.
 7. The method of claim 6, wherein the light chain variable region of the antibody has the amino acid sequence as shown in SEQ ID NO. 6 and the heavy chain variable region of the antibody has the amino acid sequence as shown in SEQ ID NO:
 7. 8. The method of claim 1, wherein the light chain constant region of the antibody has the amino acid sequence of the constant region of kappa chain.
 9. The method of claim 1, wherein the heavy chain constant region of the antibody has the amino acid sequence of the constant region of IgG2a.
 10. The method of claim 8, wherein the light chain constant region of the antibody has the amino acid sequence set forth in any one of SEQ ID NOs: 8, 10 to 12 and the heavy chain constant region of the antibody has the amino acid sequence set forth in SEQ ID NO: 9 or
 13. 11. The method of claim 1, wherein the antibody has a four-chain structure comprising two light chains and two heavy chains.
 12. The method of claim 1, wherein the cancer and/or infection is a neoplastic disease, leukemia, Johne's disease, anaplasmosis, bacterial mastitis, mycotic mastitis, a mycoplasma infection, tuberculosis, Theileria orientalis infection, cryptosporidiosis, coccidiosis, trypanosomiasis or leishmaniasis.
 13. The method of claim 12, wherein the mycoplasma infection is mycoplasma mastitis or mycoplasma pneumonia. 